miR-181c对耐顺铂鼻咽癌细胞凋亡的作用及其机制

Effect of miR-181c on Cisplatin-resistant Nasopharyngeal Carcinoma Cell Apoptosis and Its Mechanism

目的:探讨鼻咽癌细胞HNE1/DDP及HNE1中miR-181c的表达差别,分析miR-181c对耐顺铂鼻咽癌细胞凋亡的作用及其机制。方法:本次实验共分为低剂量HNE1/DDP组和高剂量HNE1组,采用RT-PCR方式检测转染之后的miR-181c在2株细胞之中的表达情况;使用MTT法检测转染的模拟物miR-181c mimics以及抑制物inhibitors对于2株细胞增殖抑制的作用。同时利用PI单染细胞对转染的miR-181c模拟物进行检验。结果:(1)HNE1/DDP细胞相对于细胞HNE1,抗凋亡蛋白MRP、Bcl-2表达提升,促凋亡蛋白Bax、Bim表达降低。(2)低剂量HNE1/DDP组内,转染miR-181c模拟物使miR-181c含量提升至(9.04±0.51),同时高剂量HNE1组内,转染miR-181c阻抑剂使miR-181c含量降低至(0.18±0.01)。转染48 h后,HNE1/DDP细胞株内miR-181c含量提升至(32.42±0.17),且HNE1细胞株内miR-181c含量减少。同均数相比,miR-181c的变异程度更高。(3)经过PI单染实验的细胞,低剂量HNE1/DDP组转染的miR-181c模拟物表达上升至(20.34±0.31),然而转染miR-181c抑制剂的HNE1细胞却由于DDP诱导的凋亡而降低至(15.02±0.12)。结论:HNE1/DDP较亲本细胞HNE1具有较强的耐药性。HNE1与HNE1/DDP细胞中的miR-181c的表达能力不一样,同时过表达的miR-181c会加速HNE1/DDP细胞死亡。

Objective: To investigate the expression difference of miR-181 c in nasopharyngeal carcinoma cells HNE1/DDP and HNE1, and to analyze the effect and mechanism of miR-181 c on cisplatin-resistant nasopharyngeal carcinoma cells. Methods: This experiment was divided into a low-dose HNE1/DDP group and a high-dose HNE1 group. The expression of miR-181 c in the two cells was detected by RT-PCR. MTT assay was used to detect the effects of miR-181 c mimics and inhibitors on the proliferation inhibition of the two cells. At the same time, PI-stained cells were used to test the transfected miR-181 c mimic. Results:(1)Compared with HNE1, the expression of anti-apoptotic proteins MRP and Bcl-2 in HNE1/DDP cells increased, while the expression of pro-apoptotic proteins Bax and Bim decreased.(2)In the low-dose HNE1/DDP group, the simulation of transfection of miR-181 c increased the miR-181 c content to(9.04±0.51), while in the high-dose HNE1 group, the transfection of miR-181 c suppressor caused miR-181 c. The content was reduced to(0.18±0.01). 48 hours after transfection, the miR-181 c content in HNE1/DDP cell lines increased to(32.42±0.17), and the miR-181 c content in HNE1 cell lines decreased. Compared with the mean, miR-181 c had a higher degree of variation.(3)After PI single-staining experiments, the expression of miR-181 c mimics in the low-dose HNE1/DDP group increased to(20.34±0.31), whereas HNE1 cells transfected with miR-181 c inhibitors were induced by DDP. Apoptosis was reduced to(15.02±0.12). Conclusion: HNE1/DDP has stronger drug resistance than HNE1. The expression capabilities of miR-181 c in HNE1 and HNE1/DDP cells are different, and overexpression of miR-181 c will accelerate the death of HNE1/DDP cells.