miRNA-30a-3p通过调控Caspase1介导的焦亡抑制肝癌细胞增殖和转移

MiRNA-30a-3p inhibits proliferation and metastasis of hepatocellular carcinoma cells by regulating caspase 1-mediated pyroptosis

目的探讨miRNA-30a-3p通过靶向作用Caspase1介导的焦亡，抑制肝癌细胞的增殖、侵袭和转移。方法利用qRT-PCR和免疫组化染色检测肝癌组织中miR-30a-3p和Caspase1的表达。用肝癌细胞SMMC-7721转染miR-30a-3p激动剂、抑制剂及Caspase1特异性抑制剂。Westernblot检测EMT（epithelialmesenchymaltransition）相关蛋白（N-cadherin、vimentin、snail、MMP．2）和Caspase1、IL-18和IL-1β蛋白表达量。平板克隆、CCK-8和Transwell实验检测肝癌细胞增殖、转移能力。结果肝癌组织中Caspase1高表达（t=17．54，P〈0．05）。过表达miR-30a-3p能抑制肝癌细胞增殖和转移能力，而抑制miR-30a-1β表达可提高肝癌细胞增殖和转移能力。过表达miR030a-3p可降低Caspase1表达（t=12．73，P〈0．05），抑制焦亡诱导，进而抑制炎症介质IL-18（t=3．51，P〈0．05）和IL-1β（t=7．32，P〈0．05）表达。抑制miRNA-30a-3p表达，活性肝癌细胞数量增加（Ft=9．57，P〈0．05）；而miRNA-30a-3p和Caspase1联合抑制活性肝癌细胞数量则减少（-=10．66，P〈0．05）。结论Caspase1在肝癌组织中高表达，miRNA-30a-3p通过作用Caspase1通路以调节细胞焦亡，进而抑制肝癌细胞增殖侵袭和转移。

Objective To investigate the rol-e of miRNA-30a-3p on inhibiting the proliferation, invasion and metastasis of HCC cells by targeting Caspase 1 involved in pyroptosis. Methods The qRT- PCR and immunohistochemical staining were used to detect the expressions of miRNA-30a-3p and Caspase 1 in HCC cells. SMMC-7721 cells were transfected with miR-30a-3p agonists, inhibitors and Caspase 1- specific inhibitors. Western blot was obtained to detect the expression of EMT-related proteins （ N-cadherin, vimentin, snail, MMP-2） and Caspase 1, IL-18 and IL--1β. Plate clone assay, CCK-8 kit and Transwell were carried out to detect the proliferation and immigration of HCC ceils. Results Caspase 1 was highly expressed （ t = 17. 54, P 〈 0. 05 ） in HCC tissues. Overexpression of miR-30a-3p inhibited HCC cells proliferation and metastasis, while miR-30a-3p inhibition increased the proliferation and metastasis of HCC cells. Overexpression of miR-30a-3p decreased the expression of Caspase 1 （ t = 12.73, P 〈 0. 05 ） and inhibited the induction of pyroptosis, inhibiting the expression of IL-18 （ t = 7.32, P 〈 0. 05 ） and IL-1β （t = 7. 32, P 〈 0. 05 ）. When miRNA-30a-3p was inhibited, the cell viability of HCC was increased （F1 = 9. 57 ,P 〈0. 05 ）. When miRNA-30a-3p and Caspase 1 were inhibited together, the cell viability of HCC decreased （F2 = 10. 66,P 〈 0. 05）. Conclusion MiRNA-30a-3p regulate cell pyroptosis through Caspase 1 pathway, inhibiting the proliferation, invasion and metastasis of HCC cells.