中国肿瘤外科杂志2024，Vol.16Issue(2)：170-177,8.DOI:10.3969/j.issn.1674-4136.2024.02.013
GM130通过14-3-3ζ对卵巢癌细胞增殖的影响及机制研究
Effect of GM130 on the proliferation of ovarian cancer cells via 14-3-3ζ and mechanism study
摘要
Abstract
Objective To investigate the effects and possible mechanisms of Golgi matrix protein 130(GM130)on ovarian cancer cell proliferation via 14-3-3ζ.Methods The expression of GM130 and 14-3-3ζ in 20 normal ovarian epithelial tissues and 42 ovarian epithelial malignant tumours was detected by immunohistochemistry,and their correlation was analyzed.Western blot and RT-PCR were used to detect the expression of GM130 and 14-3-3ζ proteins and their mRNAs in ovarian cancer SKOV3/A2780 cell lines,and the highly expressed cell lines were screened out.Immunofluorescence staining was used to detect the co-expression of GM130 and 14-3-3ζ in ovarian cancer cells.The designed recombinant expression plasmid shRNA-GM130 fragment was stably transfected into ovarian cancer cells,and Western blot and RT-PCR were used to screen out the best fragment to decrease GM130.The cells were divided into three groups:blank group(WT),control group(NC),and low-expression group(313),and the reproductive status and cycle changes of ovarian cancer cells in each group were detected by CCK-8 and flow cytometry respectively,and the expression levels of 14-3-3ζ,GRASP65,P115 and proliferation-related proteins in each group were detected by Western blot.Results The expression levels of GM130 and 14-3-3ζ were significantly higher in ovarian malignant tumour tissues than in normal ovarian tissues(P＜0.001).In ovarian malignant tumour tissues,GM130 and14-3-3ζ showed a positive correlation(r=0.873,P＜0.001).The expression of GM130 protein and its mRNA was higher in SKOV3 cells than in A2780 cells(P＜0.05),and there was no significant difference between 14-3-3ζ protein and its mRNA when comparing SKOV3 cells and A2780 cells(P＞0.05).Therefore,SKOV3 cells were selected for subsequent experiments.Immunofluorescence co-localization staining showed that GM130 and 14-3-3ζ were mainly expressed in the cytoplasm,with higher stacking-like expression in the proximal nuclear membrane portion of the former,and less expression in the cytosol of the latter,which may be co-localized.The down-regulation of GM130 was best after transfection of the fragment of group 313. The proliferation ability of cells in the transfected plasmid 313 group was significantly reduced.The number of cells blocked in the G1 phase increased,the number of cells blocked in the G2 phase was significantly reduced,the number of cells blocked in the S phase was slightly reduced,and the relative expression levels of GM130,14-3-3ζ,P115,Cyclin A,and Cdc25A proteins were reduced,while the relative expression level of P21 protein was elevated(P＜0.05).Conclusions Inhibition of GM130 inhibits ovarian cancer cell proliferation,and the mechanism may be related to the inhibition of 14-3-3ζ expression,which affects the ovarian cancer cell cycle.关键词
卵巢癌/高尔基体基质蛋白130/14-3-3ζ/细胞增殖/细胞周期Key words
Ovarian cancer/Golgi matrix protein 130/14-3-3ζ/Cell proliferation/Cell cycle引用本文复制引用
郑依果,易永芬,罗祎..GM130通过14-3-3ζ对卵巢癌细胞增殖的影响及机制研究[J].中国肿瘤外科杂志,2024,16(2):170-177,8.基金项目
国家自然科学基金资助项目(编号:30672431) （编号:30672431）
