miR-221-3p对胰腺癌细胞凋亡的抑制作用及机制研究

Inhibitory effect of miR-221-3p on apoptosis of pancreatic cancer cells and its mechanism study

钱静 ¹石宇 ¹严晓娣 ¹卞银珠²

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作者信息

1. 南通大学附属医院放疗科,江苏 226001
2. 南京大学医学院附属盐城第一医院/盐城市第一人民医院肿瘤科
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摘要

目的:研究miR-221-3p对胰腺癌细胞凋亡的抑制作用及可能机制.方法:对人胰腺癌细胞株PATU 8988T进行miR-221-3p质粒转染,设为miR-221-3p mimics组、mimics NC组、miR-221-3p inhibitor组及inhibitor NC组,以未处理的PATU8988T细胞作为空白对照(NC)组.采用实时荧光定量PCR(qRT-PCR)验证细胞转染效率;采用流式细胞术观察miR-221-3p对胰腺癌细胞凋亡的影响;采用Western blotting法测定各组细胞中p53、磷酸酶与张力蛋白同源物(phosphatase and tensin homolog,PTEN)蛋白表达.结果:qRT-PCR结果显示,与NC组、mimics NC组及inhibitor NC组比较,miR-221-3p mimics组细胞miR-221-3p表达显著增高,miR-221-3p inhibitor组表达明显降低,差异均有统计学意义(P<0.05).流式细胞术检测结果显示,与NC组、mimics NC组及inhibitor NC组比较,miR-221-3p inhibitor组细胞凋亡明显增加,miR-221-3p mimics组细胞凋亡明显减少,差异均有统计学意义(P<0.05).Western blotting结果显示,mimics NC组和inhibitor NC组细胞中p53、PTEN蛋白表达水平比较,差异无统计学意义(P＞0.05),而miR-221-3p mimics组细胞中p53、PTEN蛋白表达水平较mimics NC组明显下降,差异均有统计学意义(P<0.05).结论:PATU 8988T胰腺癌细胞中miR-221-3p表达上调,细胞凋亡受到抑制.miR-221-3p可能通过下调p53、PTEN蛋白表达促进胰腺癌的发生发展.

Abstract

Objective:To study the inhibitory effect of miR-221-3p on apoptosis of pancreatic cancer cells and its possible mechanism.Methods:Human pancreatic cancer cell line PATU8988T was transfected with miR-221-3p plasmid and divided into the miR-221-3p mimics group,the mimics NC group,the miR-221-3p inhibitor group and the inhibitor NC group.Untreated PATU8988T cells were used as blank control(NC)group.Cell transfection efficiency was verified by real-time fluorescence quantitative PCR(qRT-PCR);flow cytometry was used to observe the effect of miR-221-3p on apoptosis of pancreatic cancer cells;Western blotting was used to determine the expression of p53 and phosphatase and tensin homolog(PTEN)proteins in each group of cells.Results:qRT-PCR results showed that compared with the NC group,the mimics NC group,and the inhibitor NC group,the expression of miR-221-3p in the miR-221-3p mimics group was significantly increased,while the expression of miR-221-3p in the miR-221-3p inhibitor group was significantly re-duced,with statistical difference(P<0.05).The results of flow cytometry showed that compared with the NC group,the mim-ics NC group,and the inhibitor NC group,the miR-221-3p inhibitor group showed a significant increase in cell apoptosis,while the miR-221-3p mimics group showed a significant decrease in cell apoptosis,with statistical difference(P<0.05).Western blotting results showed that there was no statistically significant difference in the expression levels of p53 and PTEN proteins between the mimics NC group and the inhibitor NC group(P＞0.05),while the expression levels of p53 and PTEN proteins in the miR-221-3p mimics group were significantly lower than those in the mimics NC group,and the dif-ferences were statistically significant(P<0.05).Conclusion:The expression of miR-221-3p is up-regulated in pancreat-ic cancer cells line PATU8988T,and cell apoptosis is inhibited.miR-221-3p may promote the occurrence and develop-ment of pancreatic cancer by down-regulating the expression of p53 and PTEN proteins.

关键词

胰腺癌/miR-221-3p/p53/磷酸酶与张力蛋白同源物/细胞凋亡

Key words

pancreatic cancer/miR-221-3p/p53/phosphatase and tensin homolog/cell apoptosis

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基金项目

南通市科技计划项目(JC22022003)

出版年

2024

交通医学

南通大学

交通医学

影响因子：0.496

ISSN：1006-2440

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