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目的 :研究miR-145对胃癌耐顺铂细胞株SGC7901/DDP顺铂耐药的影响。方法 :应用QRT-PCR检测miR-145在胃癌组织、细胞中的表达水平;MTT法和细胞克隆形成实验检测细胞活性与增殖能力;荧光素酶实验验证miR-145的靶基因;Western blot、免疫组化和免疫荧光实验检测相关蛋白表达;流式细胞术检测耐药细胞对顺铂诱导凋亡的影响。结果 :miR-145在胃癌组织、各种胃癌细胞株中呈低表达;在胃癌耐顺铂细胞株SGC7901/DDP中,miR-145呈低表达,IGF1R与IRS1呈高表达;上调miR-145增强SGC7901/DDP细胞对顺铂的敏感性;荧光素酶报告实验证实IGF1R与IRS1为miR-145的靶基因;上调miR-145显著降低靶蛋白表达,抑制SGC7901/DDP细胞增殖,促进顺铂诱导的凋亡。结论 :上调miR-145通过靶向IGF1R与IRS1逆转胃癌细胞对顺铂的耐药性。
Objective: To study the effect of miR-145 on cisplatin-resistant gastric cancer cell line SGC7901 / DDP. METHODS: The expression of miR-145 in gastric cancer tissues and cells was detected by QRT-PCR. The cell viability and proliferation were detected by MTT assay and cell clone formation assay. The target genes of miR-145 were verified by luciferase assay. Western blot The expression of related proteins was detected by histochemical and immunofluorescence assay. The effect of drug-resistant cells on cisplatin-induced apoptosis was detected by flow cytometry. Results: The expression of miR-145 was low in gastric cancer tissues and various gastric cancer cell lines. In gastric cancer cisplatin cell line SGC7901 / DDP, the expression of miR-145 was low, IGF1R and IRS1 were highly expressed, and miR-145 was up-regulated SGC7901 / DDP cells to cisplatin sensitivity; luciferase reporter assay confirmed that IGF1R and IRS1 miR-145 target gene; upregulation of miR-145 significantly reduces the target protein expression, inhibition of SGC7901 / DDP cell proliferation and promote cisplatin-induced Apoptosis. Conclusion: Up-regulation of miR-145 can reverse the drug resistance of gastric cancer cells to cisplatin by targeting IGF1R and IRS1.