1.Gushukang interferes with osteoclasts:activation of nuclear factor erythroid 2-related factor 2 regulates the c-Fos/NFATc1 pathway in the treatment of osteoporosis
Chengzhi HOU ; Jiatong HAN ; Guangcheng WEI ; Zechuan ZHUO ; Qiuyue LI ; Yong ZHAO ; Zhangjingze YU
Chinese Journal of Tissue Engineering Research 2025;29(2):279-285
BACKGROUND:It has been shown that Gushukang affects bone metabolism by regulating nucleotide and amino acid metabolism and immune mechanisms.Current research on the mechanism of Gushukang in the treatment of osteoporosis primarily focuses on osteoblast regulation and requires further improvement from the perspective of osteoclasts. OBJECTIVE:To investigate the mechanism by which Gushukang interferes with osteoclasts in the treatment of osteoporosis using RAW264.7 cells as the research model. METHODS:Twenty-four 8-week-old female Sprague-Dawley rats were randomly divided into four groups(n=6 per group):the three experimental groups were given 1,2 and 4 g/kg osteoporosis solution by gavage(2 times per day),and the control group was given an equal amount of distilled water by gavage(2 times per day).After 7 days of intragastric administration,aortic blood samples were extracted to collect serum samples using centrifugation,and serum samples from the same groups were combined to obtain the low-,medium-,and high-concentration Gushukang-containing and normal sera for the subsequent experiments.(1)RAW264.7 cells were cultured in six groups:normal serum was added to the control group;low,medium,and high concentration groups were added with low,medium,and high concentrations of Gushukang-containing serum,respectively;ML385,a nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor was given in the Nrf2 inhibitor group;and t-BHQ,a Nrf2 activator,was added in the Nrf2 activator group.Cell viability was detected using the cell counting kit-8 assay.(2)The 3rd generation RAW 264.7 cells were cultured and divided into five groups:the blank control group was added with normal serum,the osteoclast group was added with receptor activator of nuclear factor κB ligand(RANKL),and the low-,medium-,and high-concentration groups were added with low-,medium-,and high-concentration Gushukang-containing serum based on the addition of RANKL.Tartrate-resistant acid phosphate staining was performed after 5 days of culture.(3)RAW264.7 cells were cultured and divided into five groups:blank control group was cultured with normal serum,osteoclast group cultured with normal serum and RANKL,high concentration+osteoclast group cultured with RANKL+high concentration Gushukang-containing serum,osteoclast+Nrf2 agonist group cultured with RANKL+t-BHQ,and high concentration+osteoclast+Nrf2 inhibitor group cultured with RANKL+high concentration Gushukang-containing serum+ML385.Western blot assay and determination of reactive oxygen content were performed after 5 days of culture. RESULTS AND CONCLUSION:The cell counting kit-8 results indicated that Gushukang-containing serum,NRF2 inhibitor or agonist had no significant effect on RAW264.7 cell viability.Tartrate-resistant acid phosphate staining results demonstrated that Gushukang-containing serum exhibited a concentration-dependent inhibitory effect on osteoclast differentiation.Western blot analysis and determination of reactive oxygen species revealed that compared with the blank control group,Nrf2 protein expression was decreased in the osteoclast group(P<0.05),while c-Fos and NFATc1 protein expression and reactive oxygen species content were elevated(P<0.05);compared with the osteoclast group,Nrf2 protein expression was elevated and reactive oxygen species content was decreased in the high-concentration+osteoclast group,osteoclast+Nrf2 agonist group,and high-concentration+osteoclast+Nrf2 inhibitor group(P<0.05),while c-Fos and NFATc1 protein expression was decreased in the high concentration+osteoclast group and osteoclast+Nrf2 agonist group(P<0.05);compared with the high concentration+osteoclast group,Nrf2 protein expression was decreased(P<0.05)and reactive oxygen species content was elevated(P<0.05)in the high concentration+osteoclast+Nrf2 inhibitor group.To conclude,Gushukang reduces reactive oxygen species production by activating Nrf2,thereby inhibiting downstream of the c-Fos/NFATc1 pathway and suppressing osteoclast differentiation.
2.Analysis on the Current Status of Outcome Indicators in Randomized Controlled Trials of Manual Treatment for Knee Osteoarthritis
Jinguang GU ; Guangcheng WEI ; Yong ZHAO ; Yongli DONG ; Zechuan ZHUO ; Chengzhi HOU ; Weikai QIN
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(4):58-64
Objective To provide suggestions for the clinical efficacy evaluation method of manual therapy for knee osteoarthritis(KOA)by analyzing the current status of the use of outcome indicators in randomized controlled trials(RCTs)of manual therapy for KOA.Methods RCTs about manual treatment of KOA were retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Cochrane Library,Scopus and Web of Science from March 2014 to March 2024.Two researchers screened literature,extracted literature features,analyzed the RCT indicator domain,measurement tools,and measurement time points for manual treatment of KOA.Results A total of 134 articles were included,with a sample size of 17 659.A total of 138 outcome indicators were used to classify the outcome indicators:64 symptoms and signs(46.4%),54 physical and chemical examinations(39.1%),11 quality of life(8.0%),2 psychological status indicators(1.4%),2 safety indicators(1.4%),2 muscle strength testing indicators(1.4%)and 3 other indicators(2.2%).Among 134 articles,there were a total of 36 measurement time points,with a research time span of 1 hour to 1 year after treatment;91 articles(67.9%)used"clinical efficacy"as the outcome measure;27 articles(20.1%)were evaluated for safety and reported adverse events.Conclusion The RCT outcome indicators of manual therapy for KOA lack representativeness,integrality,objectivity and specificity.The rigorism of the outcome indicators should be improved from the aspects of representativeness,integrality,objectivity and specificity,in order to achieve a complete evaluation effect on disease research.
3.Programmed death-ligand 1 tumor proportion score in predicting the safety and efficacy of PD-1/PD-L1 antibody-based therapy in patients with advanced non-small cell lung cancer: A retrospective, multicenter, observational study.
Yuequan SHI ; Xiaoyan LIU ; Anwen LIU ; Jian FANG ; Qingwei MENG ; Cuimin DING ; Bin AI ; Yangchun GU ; Cuiying ZHANG ; Chengzhi ZHOU ; Yan WANG ; Yongjie SHUI ; Siyuan YU ; Dongming ZHANG ; Jia LIU ; Haoran ZHANG ; Qing ZHOU ; Xiaoxing GAO ; Minjiang CHEN ; Jing ZHAO ; Wei ZHONG ; Yan XU ; Mengzhao WANG
Chinese Medical Journal 2025;138(14):1730-1740
BACKGROUND:
This study aimed to investigate programmed death-ligand 1 tumor proportion score in predicting the safety and efficacy of PD-1/PD-L1 antibody-based therapy in treating patients with advanced non-small cell lung cancer (NSCLC) in a real-world setting.
METHODS:
This retrospective, multicenter, observational study enrolled adult patients who received PD-1/PD-L1 antibody-based therapy in China and met the following criteria: (1) had pathologically confirmed, unresectable stage III-IV NSCLC; (2) had a baseline PD-L1 tumor proportion score (TPS); and (3) had confirmed efficacy evaluation results after PD-1/PD-L1 treatment. Logistic regression, Kaplan-Meier analysis, and Cox regression were used to assess the progression-free survival (PFS), overall survival (OS), and immune-related adverse events (irAEs) as appropriate.
RESULTS:
A total of 409 patients, 65.0% ( n = 266) with a positive PD-L1 TPS (≥1%) and 32.8% ( n = 134) with PD-L1 TPS ≥50%, were included in this study. Cox regression confirmed that patients with a PD-L1 TPS ≥1% had significantly improved PFS (hazard ratio [HR] 0.747, 95% confidence interval [CI] 0.573-0.975, P = 0.032). A total of 160 (39.1%) patients experienced 206 irAEs, and 27 (6.6%) patients experienced 31 grade 3-5 irAEs. The organs most frequently associated with irAEs were the skin (52/409, 12.7%), thyroid (40/409, 9.8%), and lung (34/409, 8.3%). Multivariate logistic regression revealed that a PD-L1 TPS ≥1% (odds ratio [OR] 1.713, 95% CI 1.054-2.784, P = 0.030) was an independent risk factor for irAEs. Other risk factors for irAEs included pretreatment absolute lymphocyte count >2.5 × 10 9 /L (OR 3.772, 95% CI 1.377-10.329, P = 0.010) and pretreatment absolute eosinophil count >0.2 × 10 9 /L (OR 2.006, 95% CI 1.219-3.302, P = 0.006). Moreover, patients who developed irAEs demonstrated improved PFS (13.7 months vs. 8.4 months, P <0.001) and OS (28.0 months vs. 18.0 months, P = 0.007) compared with patients without irAEs.
CONCLUSIONS
A positive PD-L1 TPS (≥1%) was associated with improved PFS and an increased risk of irAEs in a real-world setting. The onset of irAEs was associated with improved PFS and OS in patients with advanced NSCLC receiving PD-1/PD-L1-based therapy.
Humans
;
Carcinoma, Non-Small-Cell Lung/metabolism*
;
Male
;
Female
;
Retrospective Studies
;
Middle Aged
;
Lung Neoplasms/metabolism*
;
Aged
;
B7-H1 Antigen/metabolism*
;
Programmed Cell Death 1 Receptor/metabolism*
;
Adult
;
Aged, 80 and over
;
Immune Checkpoint Inhibitors/therapeutic use*
4.Efficacy analysis of curative esophagectomy versus definitive chemoradiotherapy in clinical T1bN0M0 thoracic esophageal cancer
Wenxue WEI ; Wenjian YAO ; Chengzhi DING ; Zeheng MA ; Mengbo LIU ; Yijun ZHANG ; Shoulong LU ; Mingbo LIU ; Li WEI
Chinese Journal of Digestive Surgery 2025;24(10):1290-1297
Objective:To evaluate the efficacy of curative esophagectomy versus definitive chemoradiotherapy (dCRT) in patients with clinical T1bN0M0 thoracic esophageal cancer.Methods:The propensity score matching (PSM) and retrospective cohort study was conducted. The clinico-pathological data of 163 patients with clinical T1bN0M0 thoracic esophageal cancer who were admitted to Henan Provincial People′s Hospital from January 2014 to December 2020 were collected. There were 125 males and 38 females, aged (58.9±7.0)years. Of 163 patients, 124 cases undergoing curative transthoracic esophagectomy were allocated into the radical resection group, 39 cases undergoing dCRT were allocated into the dCRT group. Observation indicators:(1) PSM and compari-son of clinicopathological characteristics of patients between the two groups after matching; (2) complications in the radical resection group and treatment status in the dCRT group; (3) survival analysis; (4) analysis of factors influencing patients′ prognosis. Comparison of measurement data with normal distribution between groups was conducted using the Welch t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The Cox proportional hazard model was used for univariate and multivariate analyses. The Kaplan-Meier method was used to calculate survival rate and plot survival curve, and Log-rank test was used for survival analysis. PSM was performed using the 2∶1 nearest neighbor matching method. The caliper value was set as 0.05. Results:(1) PSM and comparison of clinicopathological charac-teristics of patients between the two groups after matching. Of the 163 patients, 117 cases were successfully matched, with 78 cases in the radical resection group and 39 cases in the dCRT group. After PSM, the elimination of tumor differentiation degree confounding bias ensured comparability. (2) Complications in the radical resection group and treatment status in the dCRT group.Among the 78 patients in the curative esophagectomy group, 22 cases developed complications within 30 days after surgery. There was no death within 30 days after surgery. Among the 39 patients in the dCRT group, 25 cases received concurrent chemoradiotherapy alone, 8 cases received induction chemo-therapy followed by concurrent chemoradiotherapy, 3 cases received sequential chemoradiotherapy, and 3 cases received radiotherapy alone. Among the 33 patients who received concurrent chemo-radiotherapy, 29 cases were treated with the XP regimen, and 4 cases with the FP regimen. Efficacy evaluation showed that 37 patients achieved complete remission, and 2 patients had residual lesions. Twenty-two patients developed treatment-related adverse reactions. (3) Survival analysis. After PSM, the follow-up duration was 58(range, 13-125)months in the radical resection group and 56(range, 10-129)months in the dCRT group. The postoperative 5-year overall survival rates were 95.7% and 97.1% in the radical resection group and dCRT group, respectively, showing no significant difference between the two groups ( χ2=0.001, P>0.05). The postoperative 5-year disease-free progression survival rates were 88.2% and 94.2% in the radical resection group and dCRT group, respectively, showing no significant difference between the two groups ( χ2=0.652, P>0.05). (4) Analysis of factors influencing patients prognosis. Age and pathological TNM stage were indepen-dent influencing factors for overall survival time in patients with clinical T1bN0M0 thoracic esophageal cancer ( hazard ratio=1.312, 2.945, 95% confidence interval as 1.042-1.711, 2.204-5.517, P<0.05). Age and pathological TNM stage were independent influencing factors for disease-free survival time in patients with clinical T1bN0M0 thoracic esophageal cancer ( hazard ratio=1.215, 3.301, 95% confidence interval as 1.012-1.699, 2.012-6.321, P<0.05). Conclusions:There is no significant difference in overall survival and disease-free survival between patients with clinical T1bN0M0 thoracic esophageal cancer undergoing curative esophagectomy and dCRT. The treatment modality is not an independent prognostic factor.
5.Analysis on the Current Status of Outcome Indicators in Randomized Controlled Trials of Manual Treatment for Knee Osteoarthritis
Jinguang GU ; Guangcheng WEI ; Yong ZHAO ; Yongli DONG ; Zechuan ZHUO ; Chengzhi HOU ; Weikai QIN
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(4):58-64
Objective To provide suggestions for the clinical efficacy evaluation method of manual therapy for knee osteoarthritis(KOA)by analyzing the current status of the use of outcome indicators in randomized controlled trials(RCTs)of manual therapy for KOA.Methods RCTs about manual treatment of KOA were retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Cochrane Library,Scopus and Web of Science from March 2014 to March 2024.Two researchers screened literature,extracted literature features,analyzed the RCT indicator domain,measurement tools,and measurement time points for manual treatment of KOA.Results A total of 134 articles were included,with a sample size of 17 659.A total of 138 outcome indicators were used to classify the outcome indicators:64 symptoms and signs(46.4%),54 physical and chemical examinations(39.1%),11 quality of life(8.0%),2 psychological status indicators(1.4%),2 safety indicators(1.4%),2 muscle strength testing indicators(1.4%)and 3 other indicators(2.2%).Among 134 articles,there were a total of 36 measurement time points,with a research time span of 1 hour to 1 year after treatment;91 articles(67.9%)used"clinical efficacy"as the outcome measure;27 articles(20.1%)were evaluated for safety and reported adverse events.Conclusion The RCT outcome indicators of manual therapy for KOA lack representativeness,integrality,objectivity and specificity.The rigorism of the outcome indicators should be improved from the aspects of representativeness,integrality,objectivity and specificity,in order to achieve a complete evaluation effect on disease research.
6.Efficacy analysis of curative esophagectomy versus definitive chemoradiotherapy in clinical T1bN0M0 thoracic esophageal cancer
Wenxue WEI ; Wenjian YAO ; Chengzhi DING ; Zeheng MA ; Mengbo LIU ; Yijun ZHANG ; Shoulong LU ; Mingbo LIU ; Li WEI
Chinese Journal of Digestive Surgery 2025;24(10):1290-1297
Objective:To evaluate the efficacy of curative esophagectomy versus definitive chemoradiotherapy (dCRT) in patients with clinical T1bN0M0 thoracic esophageal cancer.Methods:The propensity score matching (PSM) and retrospective cohort study was conducted. The clinico-pathological data of 163 patients with clinical T1bN0M0 thoracic esophageal cancer who were admitted to Henan Provincial People′s Hospital from January 2014 to December 2020 were collected. There were 125 males and 38 females, aged (58.9±7.0)years. Of 163 patients, 124 cases undergoing curative transthoracic esophagectomy were allocated into the radical resection group, 39 cases undergoing dCRT were allocated into the dCRT group. Observation indicators:(1) PSM and compari-son of clinicopathological characteristics of patients between the two groups after matching; (2) complications in the radical resection group and treatment status in the dCRT group; (3) survival analysis; (4) analysis of factors influencing patients′ prognosis. Comparison of measurement data with normal distribution between groups was conducted using the Welch t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The Cox proportional hazard model was used for univariate and multivariate analyses. The Kaplan-Meier method was used to calculate survival rate and plot survival curve, and Log-rank test was used for survival analysis. PSM was performed using the 2∶1 nearest neighbor matching method. The caliper value was set as 0.05. Results:(1) PSM and comparison of clinicopathological charac-teristics of patients between the two groups after matching. Of the 163 patients, 117 cases were successfully matched, with 78 cases in the radical resection group and 39 cases in the dCRT group. After PSM, the elimination of tumor differentiation degree confounding bias ensured comparability. (2) Complications in the radical resection group and treatment status in the dCRT group.Among the 78 patients in the curative esophagectomy group, 22 cases developed complications within 30 days after surgery. There was no death within 30 days after surgery. Among the 39 patients in the dCRT group, 25 cases received concurrent chemoradiotherapy alone, 8 cases received induction chemo-therapy followed by concurrent chemoradiotherapy, 3 cases received sequential chemoradiotherapy, and 3 cases received radiotherapy alone. Among the 33 patients who received concurrent chemo-radiotherapy, 29 cases were treated with the XP regimen, and 4 cases with the FP regimen. Efficacy evaluation showed that 37 patients achieved complete remission, and 2 patients had residual lesions. Twenty-two patients developed treatment-related adverse reactions. (3) Survival analysis. After PSM, the follow-up duration was 58(range, 13-125)months in the radical resection group and 56(range, 10-129)months in the dCRT group. The postoperative 5-year overall survival rates were 95.7% and 97.1% in the radical resection group and dCRT group, respectively, showing no significant difference between the two groups ( χ2=0.001, P>0.05). The postoperative 5-year disease-free progression survival rates were 88.2% and 94.2% in the radical resection group and dCRT group, respectively, showing no significant difference between the two groups ( χ2=0.652, P>0.05). (4) Analysis of factors influencing patients prognosis. Age and pathological TNM stage were indepen-dent influencing factors for overall survival time in patients with clinical T1bN0M0 thoracic esophageal cancer ( hazard ratio=1.312, 2.945, 95% confidence interval as 1.042-1.711, 2.204-5.517, P<0.05). Age and pathological TNM stage were independent influencing factors for disease-free survival time in patients with clinical T1bN0M0 thoracic esophageal cancer ( hazard ratio=1.215, 3.301, 95% confidence interval as 1.012-1.699, 2.012-6.321, P<0.05). Conclusions:There is no significant difference in overall survival and disease-free survival between patients with clinical T1bN0M0 thoracic esophageal cancer undergoing curative esophagectomy and dCRT. The treatment modality is not an independent prognostic factor.
7.Comparison of the efficacy of neoadjuvant chemotherapy versus neoadjuvant chemoradiotherapy in locally advanced mid-low rectal cancer
Qi SUN ; Chang LIU ; Jianqiao YE ; Wenbo HUANG ; Yuhang XU ; Chengzhi YAO ; Wei ZHAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(2):278-283
【Objective】 To investigate the differences in efficacy and long-term prognosis between locally progressive low and intermediate rectal cancer patients receiving fluorouracil-based neoadjuvant chemotherapy alone (mFOLFOX6/CapeOX) and neoadjuvant radiotherapy, and to compare the therapeutic efficacy in the two groups. 【Methods】 We retrospectively analyzed the clinicopathological data of 118 patients with locally progressive low and intermediate rectal cancer who received neoadjuvant therapy from January 2019 to December 2021 at The First Affiliated Hospital of Xi’an Jiaotong University, including gender, age, body mass index (BMI), and other clinicopathological parameters. The t-test, Mann Whitney test, chi-square test or Fisher’s exact test were used to compare the differences between the two groups of patients who received neoadjuvant chemotherapy alone or neoadjuvant radiochemotherapy in terms of short-term efficacy, lymph node manifestations and long-term prognosis, respectively. Survival rates were calculated and survival curves were plotted using the Kaplan-Meier method. 【Results】 In terms of efficacy, patients in the neoadjuvant radiotherapy group achieved better tumor regression (Z=-2.05, P=0.04) and solid tumor efficacy (Z=-2.42, P=0.015), but the difference between the two groups in terms of downstaging effect of clinical stage was not statistically significant. The number of lymph nodes detected was significantly lower in the neoadjuvant radiotherapy group (neoadjuvant chemotherapy vs. neoadjuvant radiochemotherapy, 13.19±3.83 vs. 9.55±4.00, t=5.02, P<0.001), but the two groups did not differ significantly in the number of lymph node positives and lymph node positive ratio. In terms of long-term prognosis, there was no statistically significant difference in the overall survival rate or disease-free survival rate of the two groups. 【Conclusion】 Compared with neoadjuvant chemotherapy alone, neoadjuvant radiotherapy showed better short-term efficacy in patients with locally progressive low and intermediate rectal cancer, but there was no statistically significant difference between the two treatment regimens in terms of long-term prognosis.
8.Source and intension of the"Tendon Constraining Bone"theory
Guangcheng WEI ; Weikai QIN ; Yong ZHAO ; Chengzhi HOU ; Linqiang BAN ; Zechuan ZHUO ; Jinguang GU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(6):765-772
"Tendon Constraining Bone"is an essential theory in osteology and traumatology of traditional Chinese medicine,originating from the statement in Suwen(Plain Questions)that"convergent tendon controlling bones and joints".Since the Yuan and Ming dynasties,the theory of"Tendon Constraining Bone"was formed based on the understanding of anatomical relationships and the need for pathogenesis interpretation,developed by medical practitioners such as ZHU Danxi and ZHANG Jie.The"Tendon Constraining Bone"theory summarizes the physiological connections between tendons and bones and between tendons and zang-fu organs and meridians.Tendons and bones are structurally connected and functionally related,reflected in sturdy bones and tough tendons,upright bones and soft tendons with smoothly-flowing qi and blood,all tendons being related to joints,and thews being related to bones.Tendons and bones are related by meridians and zang-fu organs,specifically reflected in the Yangming channel governing the nourishment of tendons,liver governing tendons,kidneys governing bones,Taiyang channel governing tendons,and Shaoyang channel governing bones.The abnormality of"Tendon Constraining Bone"is the general pathogenesis of various tendon and bone diseases,and it can be caused by changes in the tendon and bone structure,nourishment deficiency,or the pathogenic qi retention.The pathological manifestations of abnormal"Tendon Constraining Bone"are manifested in form and state.Abnormalities in form can manifest as tendon rupture,bone fractures,tendon dislocation,and bone dislocation,whereas abnormalities in state can manifest as tendon urgency,bone pain,tendon laxity,and bone softness.The"Tendon Constraining Bone"theory has influenced the development of treatment principles such as combining motion and quiescence,paying equal attention to bone and flesh,and combining internal and external treatment.This theory has guided the application of basic treatment method such as connecting and rectifying tendons and bones,smoothing tendons and relieving bones,and nourishing tendons and strengthening bones.Therefore,the"Tendon Constraining Bone"theory can significantly guide tendon and bone disease diagnosis and treatment.
9.circDDX17 targets miR-223-3p / RIP3 to regulate the proliferation and apoptosis of non-small cell lung cancer cells
Chengzhi DING ; Guolei WANG ; Gongqian JIANG ; Hongtao WANG ; Yuanyuan LIU ; Haoliang ZHANG ; Fang SUN ; Li WEI
Chinese Journal of Oncology 2024;46(3):239-248
Objective:To explore the molecular mechanism of circDDX17 regulating the proliferation and apoptosis of non-small cell lung cancer cells by targeting the miR-223-3p/RIP3 molecular axis.Methods:The expression levels of circDDX17, miR-223-3p, and RIP3 in human normal lung epithelial cell lines BEAS-2B and non-small cell lung cancer cells H1299, A549, and H446 were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). The plasmids of pcDNA, pcDNA-circDDX17, anti-miR-con, anti-miR-223-3p, pcDNA-circDDX17 and miR-con, pcDNA-circDDX17 and miR-223-3p mimics were transfected into H1299 cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay was used to detect the cell proliferation. Flow cytometry was used to detect the cell cycle and cell apoptosis. Plate cloning experiment was used to detect cell proliferation ability. The dual luciferase report experiment was applied to verify the targeting relationship between miR-223-3p with circDDX17 and RIP3. Western blot was used to detect the protein expression of cyclinD1, CDK2, cleaved caspase-3 and Bax.Results:The expression levels of circDDX17 and RIP3 mRNA in H1299, A549, and H446 cells were significantly reduced ( P<0.05), the expression level of miR-223-3p mRNA was significantly increased ( P<0.05) compared with BEAS-2B. The cell viability [(69.46±4.68)%], the number of cell clones (83.49±7.86), the proportion of cells in S phase [(22.52±1.41) %], the protein expression levels of cyclinD1 and CDK2 in PCDNa-CircDDX17 group were lower than those in pcDNA group [(97.54±7.72)%, 205.03±13.37, (28.69±1.49)%, respectively, P<0.05], while the percentage of G 0/G 1 phase cells [(64.45±3.56)%], apoptosis rate [(18.36±1.63)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17 group were higher than those of pcDNA group [(51.33±2.76) % and (5.21±0.54) %, respectively, P<0.05]. The viability [(72.64±5.44)%], the number of cell clones (78.16±8.23), the proportion of S-stage cells [(21.34±1.59) %], the protein expression levels of CyclinD1 and CDK2 in anti-miR-223-3p group were lower than those in anti-miR-con group [(103.47±6.25)%, 169.32±14.53, (28.43±1.26)%, respectively, P<0.05]. Percentage of G 0/G 1 phase cells [(62.86±3.28)%], apoptosis rate [(14.64±1.67)%], the protein expression levels of cleaved caspase-3 and Bax in the anti-miR-223-3p group were higher than those of anti-miR-con group [(51.33±2.71)% and (4.83±0.39)%, respectively, P<0.05]. MiR-223-3p has complementary sites with circDDX17 or RIP3. The viability [(135.45±9.28)%], the number of cell clones (174.64±10.68), the proportion of S-phase cells [(26.39±2.25)%], the protein expression levels of cyclinD1 and CDK2 in pcDNA-circDDX17+miR-223-3p group were higher than those in pcDNA-circDDX17+miR-con group [(101.56±6.68)%, 107.65±7.62, (21.64±1.72)%, P<0.05]. Percentage of G 0/G 1 phase cells [(56.64±2.76)%], apoptosis rate [(8.34±0.76)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17+miR-223-3p group were lower than those of pcDNA-circDDX17+miR-con group [(64.03±3.48)% and (15.21±1.18)%, respectively, P<0.05]. Conclusion:circDDX17 could inhibit the proliferation and induce apoptosis of non-small cell lung cancer cells via targeting the miR-223-3p / RIP3 molecular axis.
10.circDDX17 targets miR-223-3p / RIP3 to regulate the proliferation and apoptosis of non-small cell lung cancer cells
Chengzhi DING ; Guolei WANG ; Gongqian JIANG ; Hongtao WANG ; Yuanyuan LIU ; Haoliang ZHANG ; Fang SUN ; Li WEI
Chinese Journal of Oncology 2024;46(3):239-248
Objective:To explore the molecular mechanism of circDDX17 regulating the proliferation and apoptosis of non-small cell lung cancer cells by targeting the miR-223-3p/RIP3 molecular axis.Methods:The expression levels of circDDX17, miR-223-3p, and RIP3 in human normal lung epithelial cell lines BEAS-2B and non-small cell lung cancer cells H1299, A549, and H446 were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). The plasmids of pcDNA, pcDNA-circDDX17, anti-miR-con, anti-miR-223-3p, pcDNA-circDDX17 and miR-con, pcDNA-circDDX17 and miR-223-3p mimics were transfected into H1299 cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay was used to detect the cell proliferation. Flow cytometry was used to detect the cell cycle and cell apoptosis. Plate cloning experiment was used to detect cell proliferation ability. The dual luciferase report experiment was applied to verify the targeting relationship between miR-223-3p with circDDX17 and RIP3. Western blot was used to detect the protein expression of cyclinD1, CDK2, cleaved caspase-3 and Bax.Results:The expression levels of circDDX17 and RIP3 mRNA in H1299, A549, and H446 cells were significantly reduced ( P<0.05), the expression level of miR-223-3p mRNA was significantly increased ( P<0.05) compared with BEAS-2B. The cell viability [(69.46±4.68)%], the number of cell clones (83.49±7.86), the proportion of cells in S phase [(22.52±1.41) %], the protein expression levels of cyclinD1 and CDK2 in PCDNa-CircDDX17 group were lower than those in pcDNA group [(97.54±7.72)%, 205.03±13.37, (28.69±1.49)%, respectively, P<0.05], while the percentage of G 0/G 1 phase cells [(64.45±3.56)%], apoptosis rate [(18.36±1.63)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17 group were higher than those of pcDNA group [(51.33±2.76) % and (5.21±0.54) %, respectively, P<0.05]. The viability [(72.64±5.44)%], the number of cell clones (78.16±8.23), the proportion of S-stage cells [(21.34±1.59) %], the protein expression levels of CyclinD1 and CDK2 in anti-miR-223-3p group were lower than those in anti-miR-con group [(103.47±6.25)%, 169.32±14.53, (28.43±1.26)%, respectively, P<0.05]. Percentage of G 0/G 1 phase cells [(62.86±3.28)%], apoptosis rate [(14.64±1.67)%], the protein expression levels of cleaved caspase-3 and Bax in the anti-miR-223-3p group were higher than those of anti-miR-con group [(51.33±2.71)% and (4.83±0.39)%, respectively, P<0.05]. MiR-223-3p has complementary sites with circDDX17 or RIP3. The viability [(135.45±9.28)%], the number of cell clones (174.64±10.68), the proportion of S-phase cells [(26.39±2.25)%], the protein expression levels of cyclinD1 and CDK2 in pcDNA-circDDX17+miR-223-3p group were higher than those in pcDNA-circDDX17+miR-con group [(101.56±6.68)%, 107.65±7.62, (21.64±1.72)%, P<0.05]. Percentage of G 0/G 1 phase cells [(56.64±2.76)%], apoptosis rate [(8.34±0.76)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17+miR-223-3p group were lower than those of pcDNA-circDDX17+miR-con group [(64.03±3.48)% and (15.21±1.18)%, respectively, P<0.05]. Conclusion:circDDX17 could inhibit the proliferation and induce apoptosis of non-small cell lung cancer cells via targeting the miR-223-3p / RIP3 molecular axis.

Result Analysis
Print
Save
E-mail