lncRNA Linc01296 对卵巢癌细胞OVCAR-3 顺铂耐药的影响及其与临床病理特征的关系

摘要] 目的：探究lncRNA Linc01296 对卵巢癌细胞顺铂（cisplatin，DDP）抵抗的影响及其作用机制。方法：收集2017 年10月至2018 年10 月四川省人民医院妇科卵巢癌组织样本53 例、交界性卵巢肿瘤样本22 例及良性卵巢肿块组织样本17 例，qPCR检测3 种样本Linc01296 mRNA表达的差异，并分析其与患者临床病理特征的关系。培养人卵巢癌细胞系OVCAR-3（OVCAR-3 Control）及其DDP耐药细胞系（OVCAR-3 DR），慢病毒转染表达靶向Linc01296 siRNA 的载体及对照随机靶向序列载体于OVCAR-3 DR细胞中，作为siLinc01296 和siControl 组，CCK-8 实验检测各组细胞系DDP半抑制浓度（IC50）及耐药指数，qPCR 检测OVCAR-3 Control、DR、DR siControl 及siLinc01296 组细胞系中Linc01296 及肿瘤干细胞相关基因Oct-4 及Sox-2 的mRNA表达水平，WB检测各组细胞系Oct-4 及Sox-2 的蛋白表达水平。结果：OVCAR-3 DR组细胞DDP IC50及耐药指数显著高于OVCAR-3 Control 组（均P<0.01），siLinc01296 组细胞DDP IC50及耐药指数显著低于DR siControl 组（P<0.01），且显著高于OVCAR-3 Control组（P<0.01）；DR 组细胞Linc01296、Oct-4 及Sox-2 mRNA 及蛋白表达均高于OVCAR-3 Control 组（均P<0.01），siLinc01296 组Linc01296、Oct-4 及Sox-2 mRNA及蛋白表达显著低于DR siControl 组（均P<0.01），而Oct-4 及Sox-2 mRNA及蛋白均高于OVCAR-3 Control 组（均P<0.01）；Linc01296 在卵巢癌组织中的表达显著高于交界性卵巢肿瘤及良性卵巢组织（均P<0.01），且与患者淋巴结转移及临床分期呈正相关（均P<0.05）。结论：Linc01296 可通过提高卵巢癌肿瘤干细胞标志物的表达促进细胞DDP抵抗的发生，且高表达于卵巢癌组织中，与患者病情进展密切相关。

Effect of long non-coding RNA Linc01296 on cisplatin resistance of ovarian cancer OVCAR-3 cells and its correlation to clinicopathological characteristics

Abstract] Objective: To investigate the effect of long non-coding RNA (lncRNA) Linc01296 on cisplatin (DDP) resistance in ovarian cancer cells and the mechanism. Methods: Fifty-three cases of ovarian cancer tissues, 22 cases of borderline ovarian tumor tissues and 17 cases of benign ovarian mass tissues were collected from Department of Gynecology, Sichuan Provincial People''s Hospital during October 2017 to October 2018. The differential expression of Linc01296 mRNA in above tissue samples was detected by qPCR, and its relationship with clinicopathological data was analyzed. Human ovarian cancer cell line OVCAR-3 (OVCAR-3 Control) and its DDPresistant cell line (OVCAR-3 DR) were cultured. Lentiviral vectors expressing siRNA that targeting Linc01296 or siRNA control were transfected into OVCAR-3 DR cells respectively, as siLinc01296 group and siControl group. CCK-8 assay was used to detect DDP semi-inhibitory concentration (IC50) and drug resistance index of each group; real-time fluorescence quantitative PCR (qPCR) was used to detect the mRNA expressions of Linc 01296 and tumor stem cell related genes (Oct-4 and Sox-2) in cell lines of OVCAR-3 control,DR, DR siControl and siLinc 01296; and the protein expressions of Oct-4 and Sox-2 in each group were detected by Wb. Results: DDP IC50 and drug resistance index in OVCAR-3 DR group was significantly higher than that in Control group (all P<0.01), and DDP IC50 and drug resistance index in siLinc01296 group was significantly lower than that in DR siControl group (all P<0.01), but significantly higher than that in Control group (all P<0.01). The mRNA and protein expressions of Linc01296, Oct-4 and Sox-2 in DR group were significantly higher than that in Control group (all P<0.01), while those expressions in siLinc01296 group were significantly lower than those in DR siControl group (P<0.01), and the mRNA and protein expressions of Oct-4 and Sox-2 in siLinc01296 group were higher than that in control group (P<0.01). The expression of Linc01296 in ovarian cancer tissues was significantly higher than that in borderline ovarian tumor tissues and benign ovarian mass tissues (P<0.01), which was positively correlated with lymph node metastasis and clinical stage (P<0.05). Conclusion: Linc01296 can promote the occurrence of DDP resistance via promoting the expression of cancer stem cell markers in ovarian cancer; Linc01296 is highly expressed in ovarian cancer tissues and closely related to disease progression of patients.