1.The mechanism of Prim-O-glucosylcimifugin in improving cholesterol metabolism in osteoarthritis chondrocytes via lncRNA NEAT1/miR-128-3p
Yanming LIN ; Haishui TU ; Shujie LAN ; Chao LI ; Shiyu LU ; Yue CHEN ; Changlong FU
Journal of Beijing University of Traditional Chinese Medicine 2025;48(1):55-67
Objective:
To investigate the mechanism of action of Prim-O-glucosylcimifugin (POG) to improve cholesterol metabolism in osteoarthritic (OA) chondrocytes based on the long noncoding RNA nuclear-enriched transcript 1 (lncRNA NEAT1)/microRNA-128-3p (miR-128-3p) pathway.
Methods:
For in vivo experiments, 60 mice were divided into the normal, sham operation, model, and POG groups using the random number table method, with 15 mice per group. The osteoarthritis mouse model was constructed using the modified Hulth method in the model and POG groups. Mice in the POG group were administered 30 mg/(kg·d)POG by gavage. The other groups were administered an equal amount of normal saline for 8 weeks. The cartilage tissue structure of mice in each group was observed using hematoxylin and eosin staining. Real-time PCR was used to detect changes in the lncRNA NEAT1 and miR-128-3p mRNA expression levels in the cartilage tissues of mice. Western blotting was used to detect the protein expressions of ATP-binding cassette transporter A1 (ABCA1), liver X receptor β (LXRβ), matrix metalloprotein-3 (MMP-3), and B-lymphoblastoma-2-associated X protein (Bax) in articular cartilage of mice. An enzyme-linked immunosorbent assay was used to measure the tumor necrosis factor-α (TNF-α) content in the synovial fluid of mice. A biochemical microplate assay was used to measure the total cholesterol level in the synovial fluid of mice. The in vitro experiments were divided into the negative control, interleukin-1β(IL-1β), IL-1β+ POG, IL-1β+ oe-lncRNA NEAT1, IL-1β+ oe-lncRNA NEAT1 + POG, IL-1β + miR-128-3p inhibition, and IL-1β+ miR-128-3p inhibition+ POG groups. An OA model was established by inducing chondrocytes with IL-1β for 24 h, and 90 mg/L of POG and miR-128-3p inhibitor(50 nmol/L) were administered for 48 h as an intervention. lncRNA NEAT1 expression in chondrocytes was detected using fluorescence in situ hybridization. A dual luciferase assay was used to detect the targeting relationship between lncRNA NEAT1 and miR-128-3p. Lentiviral plasmids overexpressing lncRNA NEAT1 were used to transfect mouse chondrocytes. Real-time PCR was used to detect the effect of lncRNA NEAT1 overexpression on the mRNA level of miR-128-3p in chondrocytes. Western blotting was used to detect ABCA1, LXRβ, MMP-3, and Bax protein expression in chondrocytes after lncRNA NEAT1 overexpression and miR-128-3p inhibition.
Results:
POG significantly reduced OA cartilage tissue damage. Compared with the model group, the lncRNA NEAT1 mRNA level decreased, whereas the miR-128-3p mRNA level increased in the cartilage tissue of the POG group (P<0.05). Compared with the model group, ABCA1 and LXRβ protein expression increased in the POG group, whereas MMP-3 and Bax protein expression decreased (P<0.05). The TNF-α levels decreased in the POG group compared to the model group (P<0.05). Compared with the model group, the total cholesterol level in the synovial fluid of the joint of mice in the POG group decreased (P<0.05). The mean fluorescence intensity of lncRNA NEAT1 in the IL-1β+ POG group decreased compared with the IL-1β group (P<0.05). The relative luciferase activity in the miR-128-3p mimics group bound to the lncRNA NEAT1-WT plasmid decreased compared with the miR-128-3p negative control group (P<0.05). The lncRNA NEAT1 mRNA levels decreased, whereas the miR-128-3p mRNA levels increased in the IL-1β+ oe-lncRNA NEAT1 + POG group compared with the IL-1β+ oe-lncRNA NEAT1 group (P<0.05). Compared with the IL-1β+ POG group, ABCA1 and LXRβ protein expression decreased, whereas MMP-3 and Bax protein expression increased (P<0.05).
Conclusion
POG mediates lncRNA NEAT1/miR-128-3p to improve cholesterol metabolism in OA chondrocytes.
2.Efficacy of 450 nm blue laser semiconductor treatment machine with submucosal injection of glycerol and fructose saline in bladder tumor endoscopic submucosal dissection
Fanzhuo TU ; Yan HU ; Xianyan CHEN ; Chao MAN ; Yongwei ZHAO
Journal of Modern Urology 2025;30(1):43-47
[Objective] To explore the safety and efficacy of submucosal injection of glycerol and fructose saline in bladder tumor endoscopic submucosal dissection (BT-ESD) with a 450 nm blue laser semiconductor treatment machine in the treatment of non-muscular invasive bladder cancer (NMIBC). [Methods] Clinical data of 20 patients with bladder tumor treated with submucosal injection of glycerol and fructose saline BT-ESD via a 450 nm blue laser semiconductor treatment machine at our hospital during Nov.2023 and Apr.2024 were retrospectively analyzed.The patients included 13 males and 7 females, aged (64.95±6.89) years, tumor diameter (1.02±0.24) cm, and preoperative hemoglobin (Hb) (130.55±4.36) g/L.The operation time, Hb the next day after operation, positive rate of tumor basal biopsy, postoperative bladder irrigation time, catheter indwelling time, postoperative hospital stay, complications and recurrence rate were recorded. [Results] All operations were successful, and 28 lesions were removed.The postoperative pathological results confirmed that all cases were NMIBC, and the basal mucosa biopsies were negative.The operation time was (9.40±3.14) min, postoperative Hb (130.15±4.59) g/L, bladder irrigation time (17.70±1.34) h, catheter indwelling time 1 day, and postoperative hospital stay 1 day.There were no complications such as obturator nerve reflex, bladder perforation.There were no need for blood transfusion, no conversion to open surgery, and no secondary bleeding after operation.No tumor recurrence was found during 6-month postoperative follow up. [Conclusion] Submucosal injection of glycerol and fructose saline BT-ESD with a 450 nm blue laser semiconductor treatment machine is a safe and effective method for the treatment of NMIBC.It has advantages of whole enucleation of bladder tumor, clear basal layer, small amount of bleeding, short operation time and few complications.It can be used as a day surgery and is worthy of clinical promotion.
3.Quality Evaluation and Analysis of Key Problems in Rukuaixiao Preparations
Rong CHEN ; Chao ZHANG ; Shuisheng ZHONG ; Wei CHEN ; Enyun TU ; Yaqiong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):146-155
ObjectiveBased on multi-index and multi-method detection, the quality of Rukuaixiao preparations was systematically evaluated from the perspective of characteristic components, and the existing problems were analyzed. MethodsLiquid chromatography-mass spectrometry(LC-MS) for the determination of 16 characteristic components was established to evaluate the quality of 129 batches of Rukuaixiao preparations. High performance liquid chromatography(HPLC) was established to determine the contents of salvianolic acids and tanshinones, investigate the rationality of quality control index of Salviae Miltiorrhizae Radix et Rhizoma in the standard for Rukuaixiao preparations in the 2020 edition of Pharmacopoeia of the People's Republic of China(hereinafter referred to as Chinese Pharmacopoeia) (volume Ⅰ), and trace the causes of significant difference among different batches. The processing and different extraction methods of Vaccariae Semen were tested, analyzing the impact of formulation changes across different editions of Chinese Pharmacopoeia. The LC-MS was established for determining the changes in the ratio of toosendanin and isotoosendanin after water extraction of Toosendan Fructus. The contents of active components in different parts of Gleditsiae Spina were determined to identify the reason of the low contents of characteristic components in some enterprises. ResultsBased on the comprehensive analysis of the samples from different dosage forms, the contents of characteristic components of Vaccariae Semen and Gleditsiae Spina in tablets from manufacturer B and granules from manufacturer D were significantly higher than those in tablets from manufacturer A, and tablets and capsules from manufacturer C. The contents of tanshinones in some batches of products from manufacturer A were abnormally high, potentially linked to the use of 70% ethanol reflux during extraction of Salviae Miltiorrhizae Radix et Rhizoma. All samples from manufacturer C exhibited abnormally high proportions of salvianic acid A and 3,4-dihydroxybenzaldehyde(salvianolic acid degradation products) to the total seven phenolic acids, indicating a risk of blindly pursuing tanshinol content and neglecting more effective components. The extraction rate of the characteristic components from Vaccariae Semen by decocting with water was obviously higher than that by reflux extraction with 70% ethanol. However, using the stir-fried Vaccariae Semen could reduce the loss of ingredients. The content ratio of toosendanin and isotoosendanin decreased from the crude herb to the prepared medicine when Toosendan Fructus was prepared by water decoction. The reason for the low component content of Gleditsiae Spina may be attributed to manufacturers using excessive non-medicinal parts in their formulations. ConclusionIt is suggested that enterprises should ensure the quality of raw material inputs, especially those without quality-control items in the standard, reduce the use of non-medicinal parts, and prohibit arbitrary alterations to manufacturing methods or processes. It is also recommended that Vaccariae Semen in Rukuaixiao capsules and granules should be changed to the stir-fried processed products. Isotoosendanin should be taken into consideration in the drug supervision when Toosendan Fructus is prepared into medicine by water decoction. Salvianolic acid B should be set as a quality control index for Salviae Miltiorrhizae Radix et Rhizoma when revising the pharmacopoeia standard of Rukuaixiao preparations.
4.Quality Evaluation and Analysis of Key Problems in Rukuaixiao Preparations
Rong CHEN ; Chao ZHANG ; Shuisheng ZHONG ; Wei CHEN ; Enyun TU ; Yaqiong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):146-155
ObjectiveBased on multi-index and multi-method detection, the quality of Rukuaixiao preparations was systematically evaluated from the perspective of characteristic components, and the existing problems were analyzed. MethodsLiquid chromatography-mass spectrometry(LC-MS) for the determination of 16 characteristic components was established to evaluate the quality of 129 batches of Rukuaixiao preparations. High performance liquid chromatography(HPLC) was established to determine the contents of salvianolic acids and tanshinones, investigate the rationality of quality control index of Salviae Miltiorrhizae Radix et Rhizoma in the standard for Rukuaixiao preparations in the 2020 edition of Pharmacopoeia of the People's Republic of China(hereinafter referred to as Chinese Pharmacopoeia) (volume Ⅰ), and trace the causes of significant difference among different batches. The processing and different extraction methods of Vaccariae Semen were tested, analyzing the impact of formulation changes across different editions of Chinese Pharmacopoeia. The LC-MS was established for determining the changes in the ratio of toosendanin and isotoosendanin after water extraction of Toosendan Fructus. The contents of active components in different parts of Gleditsiae Spina were determined to identify the reason of the low contents of characteristic components in some enterprises. ResultsBased on the comprehensive analysis of the samples from different dosage forms, the contents of characteristic components of Vaccariae Semen and Gleditsiae Spina in tablets from manufacturer B and granules from manufacturer D were significantly higher than those in tablets from manufacturer A, and tablets and capsules from manufacturer C. The contents of tanshinones in some batches of products from manufacturer A were abnormally high, potentially linked to the use of 70% ethanol reflux during extraction of Salviae Miltiorrhizae Radix et Rhizoma. All samples from manufacturer C exhibited abnormally high proportions of salvianic acid A and 3,4-dihydroxybenzaldehyde(salvianolic acid degradation products) to the total seven phenolic acids, indicating a risk of blindly pursuing tanshinol content and neglecting more effective components. The extraction rate of the characteristic components from Vaccariae Semen by decocting with water was obviously higher than that by reflux extraction with 70% ethanol. However, using the stir-fried Vaccariae Semen could reduce the loss of ingredients. The content ratio of toosendanin and isotoosendanin decreased from the crude herb to the prepared medicine when Toosendan Fructus was prepared by water decoction. The reason for the low component content of Gleditsiae Spina may be attributed to manufacturers using excessive non-medicinal parts in their formulations. ConclusionIt is suggested that enterprises should ensure the quality of raw material inputs, especially those without quality-control items in the standard, reduce the use of non-medicinal parts, and prohibit arbitrary alterations to manufacturing methods or processes. It is also recommended that Vaccariae Semen in Rukuaixiao capsules and granules should be changed to the stir-fried processed products. Isotoosendanin should be taken into consideration in the drug supervision when Toosendan Fructus is prepared into medicine by water decoction. Salvianolic acid B should be set as a quality control index for Salviae Miltiorrhizae Radix et Rhizoma when revising the pharmacopoeia standard of Rukuaixiao preparations.
5.Effect of the impaction of posterior wall on the prognosis following open reduction and internal fixation for fractures of acetabular posterior wall
Yufeng GE ; Feng GAO ; Chao TU ; Gang LIU ; Minghui YANG ; Xu SUN ; Zhelun TAN ; Yimin CHEN ; Weidong PENG ; Xinbao WU
Chinese Journal of Orthopaedic Trauma 2024;26(3):188-193
Objective:To evaluate the effect of the impaction of posterior wall on the prognosis following open reduction and internal fixation for fractures of acetabular posterior wall.Methods:A retrospective study was conducted to analyze the data from the 83 patients with fracture of acetabular posterior wall who had been consecutively treated by open reduction and internal fixation at Department of Orthopaedics and Traumatology, Beijing Jishuitan Hospital from January 2017 to December 2020. The patients were divided into 2 groups based on involvement of posterior wall impaction. In the impaction group of 33 cases, there were 26 males and 7 females with an age of (47.4±11.6) years; in the non-impaction group of 50 cases, there were 43 males and 7 females with an age of (41.3±12.0) years. The quality of postoperative fracture reduction, the function of the affected hip at the last follow-up, and the complication rate during follow-up were compared between the 2 groups. Multifactorial binary logistic regression and age subgroups were used to analyze the effects of posterior wall impaction on functional outcomes.Results:The age, rate of associated injuries in other body parts, and rate of posterior wall comminution in the impaction group were significantly higher than those in the non-impaction group ( P<0.05), but there was no statistically significant difference in other general data of patients between the 2 groups ( P>0.05). All patients were followed up for (44.5±13.3) months after surgery. The rate of anatomical reduction in the non-impaction group (96.0%, 48/50) was significantly higher than that in the impaction group (57.6%, 19/33) ( P<0.05), and the good and excellent rate by the modified Merle d'Aubigné & Postel scale at the last follow-up in the non-impaction group (84.0%, 42/50) was significantly higher than that in the impaction group (51.5%, 17/33) ( P<0.05). There was no significant difference in the incidence of complications between the 2 groups ( P>0.05). After adjusting for age and gender, the difference in hip function was still significantly different between the 2 groups ( OR=0.23, 95% CI: 0.06 to 0.79, P=0.020). The effect of posterior wall impaction on functional outcomes was statistically significant in patients aged ≥50 years ( P=0.008), whereas the difference was not statistically significant in patients aged <50 years ( P=0.194). Conclusions:Compared with non-impaction ones, acetabular fractures of posterior wall impaction tend to lead to poorer quality of reduction, which in turn affects the postoperative recovery of hip joint function. The impact of impaction fractures on functional recovery is more significant in patients aged 50 years and above.
6.Efficacy of"ditching and ridge removal"with 450 nm semiconductor blue laser in the treatment of large volume benign prostatic hyperplasia
Fanzhuo TU ; Xianyan CHEN ; Yaping QU ; Song LI ; Yan HU ; Chao MAN ; Changguan GE ; Yongwei ZHAO
Journal of Modern Urology 2024;29(5):435-439
Objective To explore the efficacy and safety of"ditching and ridge removal"with 450 nm semiconductor blue laser in the treatment of large volume benign prostatic hyperplasia(BPH),in order to promote the clinical application of this method.Methods A retrospective study was conducted on 30 patients with large volume BPH treated with"ditching and ridge removal"with 450 nm semiconductor blue laser in Yingsheng Branch of Tai'an Central Hospital during Sep.and Dec.2023.The laser operation time,level of hemoglobin before and after operation,bladder irrigation time after operation,urinary catheter indwelling time,postoperative hospital stay,and intraoperative and postoperative complications were recorded.The changes of international prostate symptom score(IPSS),quality of life scale(QoL)score,maximum urinary flow rate(Qmax)and post-void residual volume(PVR)were compared before and 1 month after operation.Results The volume of prostate was(104.5±14.52)mL,the laser operation time was(20.13±2.98)min,and the bladder irrigation time was(20.27±2.56)h.The catheter was removed in all patients 2 days after operation,and all patients were discharged 3 days after operation.One month after operation,the IPSS,QoL,Qmax and PVR were significantly improved as compared with those before operation(P<0.05).No complications occurred during the follow-up.Conclusion"Ditching and ridge removal"with 450 nm semiconductor blue laser is a new,safe and effective method in the treatment of large volume BPH.
7.Comparison of clinical efficacy of transurethral blue laser vaporization of prostate and transurethral resection of prostate in the treatment of benign prostatic hyperplasia
Fanzhuo TU ; Yan HU ; Xianyan CHEN ; Zunjie HU ; Chao MAN ; Yongwei ZHAO
Journal of Modern Urology 2024;29(9):803-808
Objective To compare the clinical efficacy and safety of transurethral blue laser vaporization of prostate and transurethral resection of prostate(TURP)in the treatment of benign prostatic hyperplasia(BPH).Methods The clinical data of 142 BPH patients undergoing surgery in our hospital during Jan.and Aug.2023 were retrospectively analyzed.The patients were divided into blue laser group(n=72)and TURP group(n=70).The operation time,hemoglobin(Hb)decrease,bladder irrigation time,catheter indwelling time and postoperative hospital stay were compared between the two groups.The changes of international prostate symptom score(IPSS),quality of life score(QoL),maximum urinary flow rate(Qmax),postvoid residual volume(PVR),international index of erectile function-5(IIEF-5)score and complications before and 6 months after operation were also compared.Results Six months after operation,the IPSS,QoL,Qmax and PVR were significantly improved in both groups(P<0.05),but there were no significant differences between the two groups(P>0.05).Compared with the TURP group,the blue laser group had shorter operation time[(47.04±4.03)min vs.(25.46±5.84)min],shorter postoperative bladder irrigation time[(54.17±4.99)h vs.(21.18±4.11)h],shorter catheter indwelling time[5(5,6)d vs.2(2,2)d],shorter postoperative hospital stay[7(7,8)d vs.4(3,4)d],less Hb decrease[(9.61±2.16)g/L vs.(4.31±2.34)g/L],and lower incidence of complications(51.4%vs.4.2%),with significant differences(P<0.05).Six months after surgery,the blue laser group had higher IIEF-5 score than that the TURP group[(20.06±2.13)vs.(18.93±1.85)],with significant difference(P<0.05).Conclusion Both transurethral blue laser vaporization of prostate and TURP can effectively treat BPH,but the former is more effective,for it has advantages of short recovery time,few complications and small impact on the sexual function of patients.
8.Prognostic and clinical value of circPRKCI expression in diverse human cancers
Zhongyue LIU ; Xiaolei REN ; Zhimin YANG ; Lin MEI ; Wenyi LI ; Chao TU ; Zhihong LI
Chinese Medical Journal 2024;137(2):152-161
Background::Highly expressed in various human cancers, circular RNA Protein Kinase C Iota (circPRKCI) has been reported to play an important role in cancer development and progression. Herein, we sought to reveal the prognostic and clinical value of circPRKCI expression in diverse human cancers.Methods::We searched the Pubmed, Web of Science, and the Cochrane Library databases from inception until May 16, 2021. The relationship between circPRKCI expression and cancer patients’ survival, including overall survival (OS) and disease-free survival (DFS), was assessed by pooled hazard ratios (HR) with corresponding 95% confidence interval (CI). The correlation between circPRKCI expression and clinical outcomes was evaluated using odds ratios (OR) with corresponding 95% CI. The data were analyzed by STATA software (version 12.0) or Review Manager (RevMan 5.3).Results::A total of 15 studies with 1109 patients were incorporated into our meta-analysis. The results demonstrated that high circPRKCI expression was significantly related to poor OS (HR = 1.96, 95% CI: 1.61, 2.39, P <0.001) when compared with low circPRKCI expression in diverse human cancers. However, elevated circPRKCI expression was not associated with DFS (HR = 1.34, 95% CI: 0.93, 1.95, P = 0.121). Furthermore, the patient with a higher circPRKCI expression was prone to have a larger tumor size, advanced clinical stage, and lymph node metastasis, but it was not significantly correlated with age, gender, and distant metastasis. Conclusion::Elevated circPRKCI expression was correlated with worse OS and unfavorable clinical features, suggesting a novel prognostic and predictive role of circPRKCI in diverse human cancers.
9.Curcumin inhibits the proliferation and migration of osteosarcoma by regulating the expression of super-enhancer-associated genes
Zhanbo OUYANG ; Haihong ZHU ; Zhongyue LIU ; Chao TU ; Jian QU ; Qiong LU ; Min XU
Journal of Central South University(Medical Sciences) 2024;49(4):541-552
Objective:Super-enhancer-associated genes may be closely related to the progression of osteosarcoma,curcumin exhibits a certain inhibitory effect on tumors such as osteosarcoma.This study aims to investigate the effects of curcumin on osteosarcoma in vitro and in vivo,and to determine whether curcumin can inhibit the progression of osteosarcoma by suppressing the expression of super-enhancer-associated genes LIM and senescent cell antigen-like-containing domain 1(LIMS1),secreted protein acidic and rich in cysteine(SPARC),and sterile alpha motif domain containing 4A(SAMD4A). Methods:Human osteosarcoma cell lines(MG63 cells or U2OS cells)were treated with 5 to 50 μmol/L curcumin for 24,48,and 72 hours,followed by the methyl thiazolyl tetrazolium(MTT)assay to detect cell viability.Cells were incubated with dimethyl sulfoxide(DMSO)or curcumin(2.5,5.0 μmol/L)for 7 days,and a colony formation assay was used to measure in vitro cell proliferation.After treatment with DMSO or curcumin(10,15 μmol/L),a scratch healing assay and a transwell migration assay were performed to evaluate cell migration ability.Real-time reverse transcription polymerase chain reaction(real-time RT-PCR)and Western blotting were used to detect mRNA and protein expression levels of LIMS1,SPARC,and SAMD4A in the cells.An osteosarcoma-bearing nude mouse model was established,and curcumin was administered via gavage for 14 days to assess the impact of curcumin on tumor volume and weight in vivo.Real-time RT-PCR was used to measure mRNA expression levels of LIMS1,SPARC,and SAMD4A in the cancer and adjacent tissues from 12 osteosarcoma patients. Results:After treating cells with different concentrations of curcumin for 24,48,and 72 hours,cell viability were all significantly decreased.Compared with the DMSO group,the colony formation rates in the 2.5 μmol/L and 5.0 μmol/L curcumin groups significantly declined(both P<0.01).The scratch healing assay showed that,compared with the DMSO group,the migration rates of cells in the 10 μmol/L and 15 μmol/L curcumin groups were significantly reduced.The exception was the 10 μmol/L curcumin group at 24 h,where the migration rate of U2OS cells did not show a statistically significant difference(P>0.05),while all other differences were statistically significant(P<0.01 or P<0.001).The transwell migration assay results showed that the number of migrating cells in the 10 μmol/L and 15 μmol/L curcumin groups was significantly lower than that in the DMSO group(both P<0.001).In the in vivo tumor-bearing mouse experiment,the curcumin group showed a reduction in tumor mass(P<0.01)and a significant reduction in tumor volume(P<0.001)compared with the control group.Compared with the DMSO group,the mRNA expression levels of LIMS1,SPARC,and SAMD4A in the 10 μmol/L and 15 μmol/L curcumin groups were significantly down-regulated(all P<0.05).Additionally,the protein expression level of LIMS1 in U2OS cells in the 10 μmol/L curcumin group was significantly lower than that in the DMSO group(P<0.05).Compared with adjacent tissues,the mRNA expression level of SPARC in osteosarcoma tissues was significantly increased(P<0.00l),while the mRNA expression levels of LIMS1 and SAMD4A did not show statistically significant differences(both P>0.05). Conclusion:Curcumin inhibits the proliferation and migration of osteosarcoma both in vitro and in vivo,which may be associated with the inactivation of super-enhancer-associated gene LIMS1.
10.Downregulation of B4GALNT1 inhibits proliferation and metastasis of osteosarcoma cells
Shuai LI ; Bing WANG ; Xia CHEN ; Chao TU ; Dan PENG ; Yuliang DAI
Journal of Central South University(Medical Sciences) 2024;49(6):870-877
Objective:Osteosarcoma is the most common malignant bone tumor in children and adolescents,characterized by a high potential for proliferation and metastasis.Patients with osteosarcoma who have distant metastases generally have a poor prognosis.Challenges in treatment include incomplete resection of tumor and chemotherapy resistance,with no effective cure currently available.Recent studies suggest that β-1,4-N-acetyl-galactosaminyltransferase 1(B4GALNT1)plays a role in the progression of various malignant tumors.However,the function of B4GALNT1 in osteosarcoma cells has not been reported.This study aims to investigate the expression of B4GALNT1 in osteosarcoma tissues compared to normal tissues and to explore its effects on the proliferation,migration,and invasion of osteosarcoma cells,thereby providing new theoretical foundations and directions for the treatment of osteosarcoma patients. Methods:Tumor tissues and corresponding normal tissue samples were collected from 16 osteosarcoma patients who underwent tumor resection at the Second Xiangya Hospital of Central South University.The patients'ages ranged from 8 to 17 years(median age 12 years).The expression of B4GALNT1 mRNA in osteosarcoma tissues,corresponding normal tissues,3 osteosarcoma cell lines(MG63,Saos-2,and U2OS),and human fetal osteoblastic cells(hFOB)was detected using real-time reverse transcription PCR(real-time RT-PCR).The effects of B4GALNT1 knockdown on the proliferation of osteosarcoma cells Saos-2 and U2OS were analyzed using cell counting kit-8(CCK-8)assays and colony formation assays.The effects of B4GALNT1 knockdown on the migration and invasion abilities of Saos-2 and U2OS cells were evaluated using Transwell migration and invasion assays.Western blotting analysis was performed to assess the impact of B4GALNT1 knockdown on the expression of epithelial-mesenchymal transition(EMT)and invasion-related proteins in Saos-2 and U2OS cells. Results:Real-time RT-PCR results showed that B4GALNT1 mRNA expression levels were significantly higher in osteosarcoma tissues and the 3 osteosarcoma cell lines compared to normal tissues and hFOB cells(all P<0.0 1).CCK-8 and colony formation assays indicated that B4GALNT1 knockdown significantly reduced the proliferation rate of osteosarcoma cells compared to the control group(all P<0.05).Transwell migration and invasion assays demonstrated that B4GALNT1 knockdown significantly decreased the number of migrating and invading osteosarcoma cells(all P<0.01).Western blotting analysis revealed that B4GALNT1 knockdown inhibited the expression of N-cadherin,Snail,Vimentin,and matrix metalloproteinase 9(MMP9)compared to the control group(all P<0.01). Conclusion:B4GALNT1 is upregulated in osteosarcoma tissues and cell lines,and its knockdown suppresses the malignant phenotype of osteosarcoma cells.B4GALNT1 may function as an oncogene in the proliferation and metastasis of osteosarcoma cells.


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