目的:探讨长链非编码RNA(lncRNA)MVIH与胃癌细胞顺铂耐药的关系及其调控机制。方法:构建胃癌顺铂耐药细胞,CCK8法检测细胞增殖能力和顺铂半数抑制浓度(IC50),并计算耐药系数;实时定量PCR(qRT-PCR)分析耐药细胞中lncRNA MVIH和let-7b-5p的表达情况;流式细胞术检测细胞凋亡率;蛋白质印迹法检测凋亡相关蛋白Bax和Caspase3的表达;通过lncLocator数据库和lncRNASNP2数据库分析lncRNA MVIH在细胞中的分布情况和可能调控的miRNA,双萤光素酶报告基因实验进行验证;生物信息学分析let-7b-5p在胃癌和癌旁组织的表达情况以及对胃癌预后的影响。结果:胃癌顺铂耐药细胞MGC803R中lncRNA MVIH的表达高于其亲本细胞MGC803(P＜0.05);在MGC803R中转染siMVIH显著增加了顺铂诱导的细胞凋亡,凋亡相关蛋白Bax和Caspase3显著上调,同时抑制了其增殖(P＜0.05);lncRNA MVIH富集在细胞质中,可以靶向调控let-7b-5p,且let-7b-5p在胃癌中低表达,并与胃癌预后呈正相关(P＜0.05);在MGC803中同时转染siMVIH和let-7b-5p抑制剂,逆转了lncRNA MVIH对细胞顺铂敏感性的调控。结论:LncRNA MVIH通过调节let-7b-5p促进胃癌细胞顺铂耐药。
Objective: To investigate the relationship between long non-coding RNA (lncRNA) MVIH and cisplatin resistance in gastric cancer cells and its detailed regulatory mechanism. Methods: The cisplatin-resistant cells of gastric cancer were constructed. The proliferation capacity and cisplatin half-inhibitory concentration (IC50) were detected by CCK8 assay, and the resistance index was calculated. Real-time quantitative PCR (qRT-PCR) was used to analyze the expressions of lncRNA MVIH and let-7b-5p in resistant cells. The apoptosis rate was detected by flow cytometry. The expressions of apoptosis-related proteins Bax and Caspase3 were detected by western blotting. lncRNA MVIH distribution in cells and possible regulated miRNAs were analyzed by lncLocator database and lncRNASNP2 database, and dual luciferase reporter gene assay was used for verification. Bioinformatics was used to analyze the expression of let-7b-5p in gastric cancer and adjacent tissues and its effect on prognosis of gastric cancer. Results: The expression of lncRNA MVIH was higher in gastric cancer cisplatin-resistant cells MGC803R than its parental cells MGC803; transfection of siMVIH in MGC803R significantly increased cisplatin-induced apoptosis, significantly upregulated apoptosis-associated proteins Bax and Caspase3, while promoting the cell proliferation. LncRNA MVIH was enriched in the cytoplasm and regulated let-7b-5p, and let-7b-5p was lowly expressed in gastric cancer and positively correlated with gastric cancer prognosis; co-transfection of siMVIH and let-7b-5p inhibitor in MGC803 reversed the regulation of cisplatin sensitivity by lncRNA MVIH. Conclusion: LncRNA MVIH promotes cisplatin resistance in gastric cancer cells by regulating let-7b-5p.
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