1.Effects of sophoranone on the biological behavior of nasopharyngeal carcinoma CNE-1 cells and MAPK signaling pathway
Chen YAO ; Dongjie YUAN ; Zheng LI ; Fangfang LI ; Zhenmin LU
China Pharmacy 2025;36(18):2279-2284
OBJECTIVE To study the effects of sophoranone (SOP) on the biological behavior of nasopharyngeal carcinoma CNE-1 cells and mitogen-activated protein kinase (MAPK) signaling pathway. METHODS CNE-1 cells were divided into blank group and SOP low-, medium- and high-concentration groups (SOP-L group, SOP-M group, SOP-H group, 25, 50 and 100 μmol/L). The number of invasive cells, the number of migratory cells, and the apoptosis rate of cells were detected. The expression levels of mitogen-activated protein kinase kinase (MEK), extracellular signal-regulated kinase 1 (ERK1), ERK2, and c-Jun N-terminal kinase (JNK) mRNA, as well as phosphorylation levels of ERK, JNK, and p38 mitogen-activated protein kinase (abbreviated as “p38”) proteins in cells were all detected. Additionally, cells were divided into blank group, SOP high-concentration group (SOP- H group, 100 μmol/L), SOP high-concentration combined with p38 inhibitor group (SOP-H+SB group, 100 μmol/L SOP+10 μmol/L SB), and SOP high-concentration combined with JNK inhibitor group (SOP-H+SP group, 100 μmol/L SOP+10 μmol/L SP). The number of invasive cells, cell migration rate, and the protein phosphorylation levels of JNK and p38 in cells, as well as the protein expression levels of matrix metalloproteinase-9(MMP-9), proliferating cell nuclear antigen Ki67, and cleaved-caspase-3 were measured. RESULTS Compared with the blank group, SOP for each concentration could significantly decrease the number of invasive cells, the number of migratory cells, and mRNA expressions of MEK, ERK1, ERK2 (except for the SOP-L group) and JNK, but increase the apoptosis rate of cells and phosphorylation levels of ERK, JNK, and p38 proteins (P<0.05). Compared with the SOP-H group, the protein phosphorylation levels of p38 and JNK, and the protein expression of cleaved-caspase-3 were decreased significantly in SOP-H+SB group and SOP-H+SP group, while the number of invasive cells, cell migration rate, and the protein expression levels of MMP-9 and Ki67 were all increased significantly (P<0.05). CONCLUSIONS SOP can inhibit the proliferation, migration and invasion of CNE-1 cells, and induce the apoptosis, the mechanisms of which may be associated with promoting the phosphorylation of proteins related to the MAPK signaling pathway.
2.Comparison of modified double-reverse traction and traditional open reduction in the treatment of tibial plateau fractures
Huankun LI ; Yanhong LI ; Dongjie HUANG ; Baijun HU ; Dawei GAO ; Yufeng WU ; Jianbang TANG ; Hongjun CHEN ; Lili SANG
Chinese Journal of Orthopaedic Trauma 2024;26(2):118-124
Objective:To compare the curative effects between modified double-reverse traction technique and traditional open reduction in the treatment of tibial plateau fractures.Methods:A retrospective study was conducted to analyze the data of 70 patients with Schatzker type Ⅳ-Ⅴ tibial plateau fracture who had undergone surgical treatment at The Third Department of Orthopaedics, Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine from January 2017 to December 2022. The patients were divided into an observation group treated with modified double-reverse traction and a control group treated with traditional open reduction. In the observation group of 37 cases, there were 20 males and 17 females with an age of (44.6±13.5) years, and 9 cases of type Ⅳ and 28 cases of type V by the Schatzker classification; in the control group of 33 cases, there were 18 males and 15 females with an age of (45.9±13.7) years, and 10 cases of type Ⅳ and 23 cases of type Ⅴ by the Schatzker classification. The 2 groups were compared in terms of operation time, length of main incision, intraoperative blood loss, Rasmussen imaging score before discharge, and knee function score of American Hospital for Special Surgery (HSS), Visual Analogue Scale (VAS), fracture healing and complications at 6 months postoperatively.Results:There was no significant difference in the preoperative general data between the 2 groups, showing group comparability ( P>0.05). All patients were followed up for (14.3±1.4) months. The observation group was significantly better than the control group in operation time [(113.9±11.4) min versus (151.82±10.37) min], length of main incision [4 (4, 5) cm versus 6 (6, 7) cm], intraoperative blood loss [30 (20, 35) mL versus 55 (50, 65) mL], VAS [0 (0, 0) point versus 0 (0, 1) points] and HSS score [(89.8±3.1) points versus (86.0±3.5) points] ( P<0.05). There were no significant differences between the 2 groups in Rasmussen imaging score before discharge, or fracture healing rate or complication rate at 6 months postoperatively ( P>0.05). Conclusion:In the treatment of Schatzker type Ⅳ-Ⅴ tibial plateau fractures, modified double-reverse traction technique is worthy of clinical application and promotion, because it is advantageous over traditional open reduction in terms of shorter operation time, smaller surgical incision, less intraoperative blood loss, less postoperative pain and better knee function.
3.Application of double mediastinal drainage tubes in elderly patients with intrathoracic anastomotic leak after thoracoscopic Ivor-Lewis surgery
Chuanfei ZHAN ; Shilin CHEN ; Xiaokang SHEN ; Dongjie FENG ; Xiaojun WANG ; Weizhong SHEN ; Feng JIANG ; Qin ZHANG ; Lin XU
Chinese Journal of Geriatrics 2024;43(1):56-59
Objective:To investigate the clinical significance of intraoperative prepositioning of dual mediastinal drains in elderly patients developing anastomotic leakage(AL)after a total endoscopic Ivor-Lewis procedure.Methods:This retrospective case-control study analyzed the clinical data of 500 elderly patients who underwent total endoscopic Ivor-Lewis surgery for esophageal or cardia cancer from January 2020 to December 2022.In the control group, one mediastinal drainage tube was placed intraoperatively, while in the study group, two mediastinal drainage tubes were placed.Both groups had a chest tube placed conventionally.The study compared the incidence of anastomotic leak(AL)at 1 month postoperatively, inflammatory indexes in patients with AL, grading of AL, rate of nasal fistula placement, incision infection, anastomotic stenosis, and incidence of hoarseness.Additionally, it compared ICU occupancy, ventilator use, and ICU length of stay between the two groups.Results:The analysis included clinical data from 455 elderly patients.Among the patients who developed AL, the study group had significantly lower peak body temperature[(39.58±1.03)℃ vs.(38.05±0.56)℃, t=4.298, P<0.05], white blood cell count[(18.63±3.35)×10 9/L vs.(14.28±2.78)×10 9/L, t=3.450, P<0.05], and C-reactive protein(CRP)levels[(154.66±41.64)mg/L vs.(122.19±31.29)mg/L, t=2.131, P<0.05]. The study group also had a significantly lower grading of AL and rate of nasal fistula placement(82.4% vs.30.0%, P<0.05). In terms of ICU indicators, the study group had a significantly lower ICU admission rate(64.7% vs.10.0%, P<0.05), shorter period of ventilator use[(6.47±8.15)days vs.(0.90±2.23)days, t=2.62, P<0.05], and shorter ICU stay[(11.70±8.89)days vs.(4.70±6.27)days, t=2.184, P<0.05]. Conclusions:Double mediastinal drainage tubes, have been found to significantly alleviate inflammation, decrease the rate of nasal fistula placement and ICU admission, and shorten the length of ICU stay in elderly patients.Therefore, they are considered safe and deserving of clinical promotion.
4.Efficacy comparison between olaparib and platinum-containing regimen for treatment of platinum-sensitive relapsed ovarian cancer patients with BRCA mutation
Jing CHEN ; Nan TANG ; Yuanyuan WU ; Yuan TIAN ; Tong LIU ; Yanli WANG ; Dongjie LI ; Runpu LI
Cancer Research and Clinic 2024;36(3):177-183
Objective:To explore the differences in tumor-specific growth factors, cellular immune function and efficacy of olaparib and platinum-containing regimen for treatment of platinum-sensitive relapsed ovarian cancer patients with BRCA mutation.Methods:A retrospective cohort study was conducted. A total of 100 platinum-sensitive relapsed BRCA-mutant ovarian cancer patients in Baoding Second Central Hospital from September 2017 to March 2020 were retrospectively selected. The clinical data of the patients were analyzed, and they were divided into the olaparib group (treated with olaparib tablets) and the platinum-containing regimen group (treated with paclitaxel and platinum drugs for 6 cycles, followed by olaparib tablets maintenance therapy), with 50 patients in each group. The clinical efficacy, tumor specific growth factor [carbohydrate antigen (CA) 125, CA199, human epididymal protein 4 (HE4)] levels, cellular immune function-related indicators [T-cell subsets (proportions of CD3 + cells and CD4 + cells), CD4 + cells/CD8 + cells ratio (CD4 +/CD8 +)], and quality of life scores before treatment and after 2, 4 and 6 cycles of treatment of the two groups were compared, as well as the safety of the two groups. The data of three years of follow-up were obtained, Kaplan-Meier method was used to analyze the progression-free survival (PFS) of patients in the two groups, and log-rank test was used for comparison between groups. Results:The age of patients in the olaparib and platinum-containing regimen groups was (53±7) years old and (56±7) years old, respectively. The differences in compositions of patients with different age, body mass index, Eastern Cooperative Oncology Group (ECOG) performance status score, primary tumor location, lesion size, pathological stage, pathological type, germline BRCA mutation, and previous chemotherapy response between the two groups were not statistically significant (all P > 0.05). The objective response rate (ORR) [58.0% (29/50) vs. 38.0% (19/50)] and disease control rate (DCR) [80.0% (40/50) vs. 56.0% (28/50)] of the olaparib group after treatment were higher than those of the platinum-containing regimen group, and the differences were statistically significant (both P < 0.05). Serum CA125, CA199 and HE4 levels were gradually decreased in both groups before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05); serum CA125, CA199 and HE4 levels in the olaparib group after 2, 4 and 6 cycles of treatment were lower than those in the platinum-containing regimen group, and the differences were statistically significant (all P < 0.05). The CD3 + cells ratio, CD4 + cell ratio and CD4 +/CD8 + in the olaparib group gradually increased before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05), while those in the platinum-containing regimen group all gradually decreased (all P < 0.05); the CD3 + cells ratio, CD4 + cells ratio and CD4 +/CD8 + in the olaparib group were higher than those in the platinum-containing regimen group after 2, 4 and 6 cycles of treatment, and the differences were statistically significant (all P < 0.05). The quality of life scores of both groups increased before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05), and the quality of life scores of the olaparib group were higher than those of the platinum-containing regimen group after 2, 4 and 6 cycles of treatment, and the differences were statistically significant (all P < 0.05). The incidence of nausea, fatigue and malaise, vomiting, anemia, and diarrhea at all levels in the olaparib group was lower than those in the platinum-containing regimen group (all P < 0.05). By follow-up for 3 years, there was no statistically significant difference in PFS between the olaparib group and the platinum-containing regimen group ( P > 0.05). Conclusions:The efficacy of olaparib treatment in platinum-sensitive relapsed ovarian cancer patients with BRCA mutation is superior to platinum-containing regimen, and it can increase the level of T cells, inhibit the expression of tumor-specific growth factors, improve the quality of life, and have a positive effect on improving the safety of treatment.
5.Study on the regularity of acupoint selection in the treatment of sepsis with acupuncture and moxibustion based on data mining technology
Yuanyuan ZENG ; Fagen HUANG ; Yaxiu HUANG ; Yonglian HUANG ; Dongjie CHEN ; Boling LI ; Ping WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(4):407-412
Objective To analyze the acupoint selection rule of acupuncture and moxibustion in the treatment of gastrointestinal dysfunction associated with sepsis,thereby providing a reference for clinical treatment.Methods A systematic search was conducted in several databases,including China national knowledge infrastructure(CNKI),China Science Periodical Database,Chinese Science and Technology Journal Database,Chinese Biomedical literature database(CBM),Wanfang Data,and VIP Database,from the inception of the databases until September 30,2023.The collected literature was organized in an Excel database and subjected to descriptive analysis.Association rule analysis and cluster analysis were performed using SPSS 26.0 and SPSS Modeler 18.0 software.Results Finally,35 acupuncture and moxibustion prescriptions in 35 articles were selected for frequency statistics.The results showed 33 acupoints were used to treat gastrointestinal dysfunction in sepsis,with an overall frequency of use of 176 acupoints.The top 10 acupoints used by acupuncture and moxibustion for the treatment of gastrointestinal dysfunction in sepsis from high to low were Zusanli,Tianshu,Shangjuxu,Zhongwan,Neiguan,Xiajuxu,Guanyuan,Qihai,Shangwan,Xiawan.The top 3 were Zusanli(16.5%),Tianshu(13.6%),Shangjuxu(11.9%).The most commonly used meridians were the Foot Yangming Stomach Meridian(52.8%),Ren Meridian(23.9%),and Foot Taiyin Spleen Meridian(8.0%).The main sites for acupoint selection were the lower limbs(44.9%)and abdomen(41.5%).The most commonly used acupoints were combined acupoints below specific acupoints(35.2%),followed by recruited acupoints(27.8%).The results of high-frequency acupoint correlation analysis showed that the acupoints with the highest comprehensive support were Zusanli,Tianshu,Zhongwan,and Shangjuxu.Cluster analysis identified 5 effective clusters(Quchi acupoint-Sanyinjiao acupoint-Neiting acupoint-Daheng acupoint-Jiaji acupoint-Yinlingquan acupoint-Shangwan acupoint-Xiawan acupoint,Guanyuan acupoint-Qihai acupoint-Neiguan acupoint-Xiajuxu acupoint,Tianshu acupoint-Shangjuxu acupoint,Zhongwan acupoint,Zusanli acupoint)and 4 commonly used acupoint combinations(Guanyuan acupoint-Qihai acupoint-Neiguan acupoint-Xiajuxu acupoint,Tianshu acupoint-Shangjuxu acupoint,Zhongwan acupoint,Zusanli acupoint).Conclusions Acupuncture treatment for gastrointestinal dysfunction in sepsis is mainly based on the treatment of"spleen and stomach",with local acupoint selection and acupoint selection along the meridian as the main approach.It emphasizes the application of specific acupoints,especially the Xiahe and Mu acupoints.
6.Effects and regulatory mechanisms of FAM3C expression on proliferation, migration, and invasion of pancreatic cancer cells
Leisheng WANG ; Yue TAO ; Shuo ZHANG ; Enhong CHEN ; Dongjie YANG ; Youzhao HE ; Yong MAO ; Hao HU
Chinese Journal of Hepatobiliary Surgery 2024;30(7):531-536
Objective:To analyze the expression of sequence similarity family 3 member C (FAM3C) in pancreatic cancer and its effects on pancreatic cancer cell proliferation, migration, invasion and the underlying mechanisms.Methods:Tissue samples from four pancreatic cancer patients undergoing surgical resection at Affiliated Hospital of Jiangnan University from July 2022 to June 2023 were collected. Expression of FAM3C was assessed in both cancer and adjacent tissues using polymerase chain reaction and Western blot techniques. High FAM3C-expressing pancreatic cancer cells PANC-1 and MIA PaCa-2 were selected for further experiments involving FAM3C-siRNA and methyltransferase-like protein 3 (METTL3)-siRNA groups, alongside a negative control group. Cell counting, Transwell assays, and co-immunoprecipitation with methylation antibodies were utilized to evaluate the proliferation, migration, invasion, and methylation status of FAM3C.Results:The relative expression of FAM3C mRNA in cancer tissues of the four patients with pancreatic cancer was higher than that in adjacent tissues (1.29±0.19 vs 0.47±0.17, t=-6.48, P=0.001). Compared with the negative control group A, the proliferation ability of the FAM3C-siRNA interference group decreased, and the number of migrating and invasive cells decreased (all P<0.05). In the negative control group B of PANC-1 cells, the relative expression of FAM3C mRNA in IgG-treated cells was lower than that in the methylated antibody-treated group (1.05±0.53 vs 30.57±1.09, t=-42.04, P=0.001). After PANC-1 cells were treated with methylated antibodies, the relative expression of FAM3C mRNA in the negative control group B was higher than that in the METTL3-siRNA interference group (30.57±1.09 vs 18.17±0.50, t=17.89, P=0.001). Compared with the negative control group B, the proliferation ability, migration and invasion cell numbers of PANC-1 cells in the METTL3-siRNA interference group were reduced (all P<0.05). The results of MIA PaCa-2 and PANC-1 cells were consistent. Conclusions:FAM3C is highly expressed in pancreatic cancer and promotes the proliferation, migration and invasion of pancreatic cancer cells. METTL3 affects FAM3C expression by methylation and proliferation, migration and invasion of pancreatic cancer cells.
7.Comparative proteomic analysis of Virology PK-15 cells infected with wild-type strain and its EP0 gene-deleted mutant strain of pseudorabies virus
Di WANG ; Dongjie CHEN ; Shengkui XU ; Fang WEI ; Hongyuan ZHAO
Journal of Veterinary Science 2024;25(4):e54-
Objective:
This study examined the function of EP0 to provide a direction for its in-depth analysis.
Methods:
In this study, the EP0-deleted PRV mutant was obtained, and Tandem Mass Tagbased proteomic analysis was used to screen the differentially expressed proteins (DEPs) quantitatively in EP0-deleted PRV- or wild-type PRV-infected porcine kidney 15 cells.
Results:
This study identified 7,391 DEPs, including 120 and 21 up-regulated and downregulated DEPs, respectively. Western blot analysis confirmed the changes in the expression of the selected proteins, such as speckled protein 100. Comprehensive analysis revealed 141 DEPs involved in various biological processes and molecular functions, such as transcription regulator activity, biological regulation, and localization.
Conclusions
and Relevance: These results holistically outlined the functions of EP0 during a PRV infection and might provide a direction for more detailed function studies of EP0 and the stimulation of lytic PRV infections.
8.Dachaihu decoction protects intestinal mucosal barrier in severe heat stroke rats by inhibiting Toll-like receptor 4/nuclear factor-κB signaling pathway
Dongjie CHEN ; Yonglian HUANG ; Xiaohong PENG ; Yaxiu HUANG ; Fagen HUANG ; Aiting ZHANG ; Ping WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2023;30(5):592-596
Objective To explore the effect of Dachaihu decoction on the Toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway and gastrointestinal mucosal barrier in rats with severe heat stroke.Methods Sixty SPF grade Sprague-Dawley(SD)male rats were divided into normal control group,model group,Dachaihu decoction standard dose group and Dachaihu decoction high dose group of 15 rats in each group.The heat stroke model was replicated in the rats at temperature(40.5±0.5)℃and humidity(65.0±2.0)%;the normal control group was not treated.From 6 hours after mold making,drug intervention was carried out in the Dachaihu decoction high dose group and the Dachaihu decoction standard dose group of 3.38 g·kg-1·d-1 and 1.69 g·kg-1·d-1,every 8 hours for 2 days.Equal amounts of normal saline were administered to the normal control group and model group.At 6,24 and 48 hours after the molding,5 mL abdominal main artery blood from 5 rats were randomly collected from each group,and the blood was obtained by enzyme-linked immunosorbent assay(ELISA)to determine the levels of tumor necrosis factor-α(TNF-α),interleukin-1(IL-1),D-lactic acid,intestinal fatty acid-binding protein(I-FABP).At the same time,the ileum tissue was retained,and the protein expression of TLR4 and NF-κB in intestinal tissue was determined by Western blotting.Some of the ileal tissue was obtained for hematoxylin-eosin(HE)staining,and the intestinal histopathological changes were observed under light microscopy.Results The normal control group of rats had no significant change,and the other three groups showed heatstroke symptoms after mold making.The overall mortality in drug group were significantly lower than that in the model group[3.3%(1/30)vs.20.0%(3/15),P<0.05].Compared with the normal control group,the serum IL-1,TNF-α,I-FABP,D-lactic acid and the protein expression levels of TLR4 and NF-κB in the model group,Dachaihu decoction standard dose group and Dachaihu decoction high dose group all increased.Compared with the model group,at 24 hours and 48 hours after molding in the Dachaihu decoction standard dose group and Dachaihu decoction high dose group,the serum IL-1,TNF-α,I-FABP,D-lactic acid and the protein expression levels of TLR4 and NF-κB significant decreased[24 hours:TNF-α(ng/L):69.20±4.32,59.37±4.31 vs.76.99±5.02,IL-1(ng/L):132.68±4.93,112.59±9.64 vs.146.75±10.12,I-FABP(mmol/L):504.35±22.23,453.37±32.38 vs.542.58±13.83,D-lactic acid(mmol/L):114.55±8.52,90.57±3.09 vs.127.87±8.37,protein expression of TLR4(A value):1.50±0.08,1.23±0.01 vs.1.86±0.08,protein expression of NF-κB(A value):1.61±0.05,1.21±0.05 vs.1.97±0.08;48 hours:TNF-α(ng/L):58.46±5.13,38.98±5.53 vs.90.21±3.02,IL-1(ng/L):119.12±4.57,84.12±5.08 vs.170.20±6.21,I-FABP(mmol/L):436.04±27.63,321.85±22.03 vs.618.79±12.31,D-lactic acid(mmol/L):87.35±6.84,70.38±4.33 vs.154.14±10.83,protein expression of TLR4(A value):1.19±0.05,1.10±0.13 vs.2.09±0.06,protein expression of NF-κB(A value):1.15±0.09,0.97±0.08 vs.2.20±0.02,all P<0.05].The expression levels of TNF-α,I-FABP,TLR4 and NF-κB protein in Dachaihu decoction high dose group decreased significantly at 24 hours and 48 hours compared with the standard dose group,however,IL-1 and D-lactic acid decreased significantly at 48 hours after molding(all P<0.05).The pathology observation showed that,compared with the model group,the intestinal mucosa villus,the lamina propria drop and haemorrhage was decreased in the Dachaihu decoction standard dose group and Dachaihu decoction high dose group.Telangiectasia was reduced and no ulcer formation was observed.Conclusion Dachaihu decoction can inhibit TLR4/NF-κB signaling pathway,reduce intestinal inflammatory response,thus reduce gastrointestinal damage,and protect the gastrointestinal mucosal barrier in rats with severe heatstroke.
9.A real-world survey of the effect of voriconazole on tacrolimus blood concentration after lung transplantation
Dongjie GUO ; Pengmei LI ; Wenqian CHEN ; Lihong LIU
China Pharmacy 2023;34(20):2519-2524
OBJECTIVE To provide a reference for the dose adjustment of tacrolimus in patients who underwent lung transplantation after combined use of voriconazole. METHODS The clinical data of lung transplantation patients who used voriconazole and tacrolimus in our hospital from January 2020 to December 2022 were collected retrospectively. The effects of voriconazole on the valley concentration, daily dose and standardized blood concentration of tacrolimus were analyzed by using SPSS 21.0 software; multiple linear regression analysis was conducted for the factors that may affect the standardized blood concentration of tacrolimus. RESULTS A total of 153 lung transplantation patients were included. After the combination of voriconazole, the average daily dose of tacrolimus decreased from 3.37 mg to 0.76 mg, and valley concentration and standardized blood concentration were increased significantly (P<0.000 1). The average daily dose of voriconazole was negatively correlated with the standardized blood drug concentration of tacrolimus (P=0.000 1,r=-0.224). The valley concentration of voriconazole was positively correlated with valley concentration (P<0.000 1,r=0.316) and standardized blood concentration (P<0.000 1,r= 0.249) of tacrolimus. After combination with voriconazole, the standardized blood drug concentration of patients who underwent single lung transplantation was significantly higher than those who underwent double lung transplantation, and the standardized blood concentration of tacrolimus after oral administration of voriconazole was significantly higher than after intravenous drip of voriconazole (P<0.05). Most liver and kidney function indicators showed no significant changes. The results of multiple factor regression analysis showed that the valley concentration of voriconazole had a significant impact on the standardized blood concentration of tacrolimus (P<0.001). CONCLUSIONS The valley concentration of voriconazole has greatest influence on the blood concentration and dose adjustment of tacrolimus, which is an independent influencing factor. In clinical practice, the dose of tacrolimus should be reduced in combination with voriconazole, and therapeutic drug monitoring should be conducted for both drugs.
10.Study on the Anti-tumor Activity of Components of Ganoderma Lucidum on the Three-dimensional Culture Model of Colorectal Cancer HCT116 Cells
PAN Haitao ; CHEN Dongjie ; ZHANG Guoliang ; HU Lingjuan ; WANG Xiaotong ; ZHONG Yi ; YANG Jihong ; LI Zhenhao
Chinese Journal of Modern Applied Pharmacy 2023;40(13):1795-1809
OBJECTIVE To investigate the anti-tumor effects of the components of Ganoderma lucidum(Gc) based on the two-dimensional(2D) and three-dimensional(3D) culture of colorectal cancer HCT116 cells. METHODS The chemical compositional of the three components was identified by UPLC-Q-TOF-MS. An in vitro 3D culture model of HCT116 cells was established by using Matrigel as the matrix material, and the effects of Gc1, Gc2, Gc3, and 5-fluorouracil(5-FU) on the proliferation of HCT116 cells in 2D and 3D culture models were evaluated, and the effects of Gc3 on cell-cycle, apoptosis, drug resistance, lipid metabolism, and 5-FU's anti-tumor activity were evaluated. Cell viability was detected by CCK-8 assay. mRNA expression level of the cells was analyzed by Real-time PCR. Proteins expression level of the cells was analyzed by Western blotting. HPLC was used to detect the content of 5-FU in cells. RESULTS A total of 76, 69, and 17 compounds were identified from Gc1, Gc2, and Gc3, respectively. Compared with 2D culture, the proliferation rate of HCT116 cells was decreased in the 3D culture model, and the expression of cell cycle-promoters CDK2, CDK4, CDK6, and fatty acid synthesizer FASN, SREBP1 were significantly down-regulated. On the contrary, the expression of cell cycle-suppressor p21, p27, and lipid droplet breakdown proteins ATGL and drug resistance gene ITGB1, CDH1, ABCB1, and ABCC1 mRNA were significantly up-regulated. Gc1, Gc2, Gc3 and 5-FU inhibited the proliferation of both 2D and 3D cultured HCT116 cells in a dose dependent manner after incubation for 48 h, and the inhibitory effect of Gc3 was significantly stronger than Gc1 and Gc2. Gc3 could not only reduce the expression of CDK2, CDK4, Bcl-xl, ATGL, and LC3B proteins, but also increase the expression of p21, p27, Bax, Cleaved caspase-3, and Cleaved PARP1 proteins, and overexpression of LC3B or ATGL attenuated Gc3-induced cytotoxicity in 3D cultured HCT116 cells. In addition, Gc3 significantly inhibited the expression of ITGB1, CDH1, ABCB1, and ABCC1 mRNA, and increased the intracellular 5-FU content, and enhanced the anti-tumor activity. CONCLUSION Gc3 significantly inhibit the proliferation of 3D-cultured HCT116 cells by inhibiting cell autophagy and lipid droplet breakdown, and enhance the anti-cancer activity of 5-FU by inhibiting the expression of ITGB1, CDH1, ABCB1, and ABCC1 mRNA.


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