摘要目的:探讨microRNA-101-3p(miR-101-3p)在卵巢癌顺铂(cisplatin，DDP)耐药中的作用及其机制。方法:购买miR-101-3p模拟物(miR-101-3p mimic)、miR-101-3p抑制剂(miR-101-3p-in)、及其各自的阴性对照RNA(miR-NC mimic和miR-NC inhibitor)。应用定量逆转录聚合酶链反应(real-time reverse transcription-PCR，qRT-PCR)和western blotting检测miR-101-3p和B淋巴瘤momlv插入区1同源物(eukaryotic translation initiation factor 4 gamma 2，EIF4G2)在卵巢癌SKOV3细胞和DDP耐药SKOV3/DDP细胞中的表达水平。用miR-101-3p模拟物和miR-101-3p模拟阴性对照转染SKOV3/DDP细胞。采用CCK-8法检测miR-101-3p模拟物转染后细胞对DDP的敏感性。流式细胞仪检测转染对细胞凋亡的影响。将EIF4G2野生型和突变型荧光素酶报告质粒与miRNA-101-3p模拟物或miRNA-101-3p NC共转染，用双荧光素酶报告系统分析荧光素酶活性。结果:CCK-8法检测结果显示，miR-101-3p在SKOV3/DDP细胞中的表达水平显著低于SKOV3细胞，EIF4G2在SKOV3/DDP细胞中的表达水平显著高于SKOV3细胞。过表达miR-101-3p能够显著提高miR-101-3p mimic组中SKOV3/DDP细胞对DDP的敏感性，其IC50值为8 μmol/L和64 μmol/L，miR-101-3p mimic可抑制SKOV3/DDP细胞活性；AO/EB实验验证过表达miR-101-3p后miR-101-3p mimic能够明显诱导SKOV3/DDP细胞凋亡。Western blotting检测结果表明，miR-101-3p mimic可显著增加B细胞淋巴瘤2(B-cell lymphoma-2，Bcl-2)、Bcl-2相关X蛋白(associated X protein，Bax)及裂解型半胱天冬酶-3的表达水平( t＝3.57、3.95、4.17， P值均<0.05)。与miR-NC mimic相比，过表达miR-101-3p后能够明显降低EIF4G2的表达( t＝4.15， P<0.05)；荧光素酶活性检测数据显示，miR-101-3p的过表达会抑制野生型EIF4G2的荧光素酶活性，但不抑制突变型EIF4G2 3’UTR的荧光素酶活性。 结论:EIF4G2是miR-101-3p的直接作用靶点，MiR-101-3p可能通过靶向调控EIF4G2，调节卵巢癌对DDP的耐药性。

abstractsObjective:To investigate the role and mechanism of microRNA-101-3p(miR-101-3p) in cisplatin (DDP) resistance of ovarian cancer.Methods:miR-101-3p mimic (miR-101-3p mimic), miR-101-3p inhibitor (miR-101-3p-in), and their respective negative control RNAs (miR NC mimic and miR NC inhibitor) have been purchased. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were applied to detect the expression levels of miR-101-3p and eukaryotic translation initiation factor 4 gamma 2 (EIF4G2) in ovarian cancer SKOV3 cells and DDP-resistant SKOV3/DDP cells. SKOV3/DDP cells were transfected with miR-101-3p mimic and miR-101-3p mimic negative control. The expression of EIF4G2 was detected by real time reverse transcription-PCR (qRT-PCR) and Western blotting, and the sensitivity of cells to DDP after transfection with miR-101-3p mimic was detected by CCK-8 assay. The effect of transfection on the apoptosis was detected via flow cytometry. EIF4G2 wild-type and mutant-type luciferase reporter plasmids were co-transfected with miRNA-101-3p mimic or miRNA-101-3p NC, and luciferase activity was analyzed by dual-luciferase reporter system.Results:The results of CCK-8 method show that the miR-101-3p expression level in SKOV3/DDP cells was significantly lower than that in SKOV3 cells, while the expression level of EIF4G2 in SKOV3/DDP cells was significantly higher than that in SKOV3 cells. Overexpression of miR-101-3p significantly increased the sensitivity of SKOV3/DDP cells to DDP in the miR-101-3p mimic group with an IC50 of 8 μmol/L and 64 μmol/L. miR-101-3p mimic could inhibit the activity of SKOV3/DDP cells. AO/EB experiment verified that miR-101-3p mimic could significantly induce apoptosis of SKOV3/DDP cells after overexpression of miR-101-3p. Western blotting showed that miR-101-3p mimic could significantly increase the expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and lysed caspase-3 ( t=3.57, 3.95, 4.17, all P values <0.05). Compared with miR-NC mimic, overexpression of miR-101-3p could significantly reduce the expression of EIF4G2 ( t=4.15, P<0.05). The luciferase activity test data showed that overexpression of miR-101-3p inhibited the luciferase activity of wild-type EIF4G2, but did not inhibit the luciferase activity of mutant EIF4G2 3 'UTR. Conclusion:EIF4G2 is the direct target of miR-101-3p. miR-101-3p may regulate ovarian cancer's sensitivity to DDP by targeted regulation on EIF4G2.