Department of Hepatopancreatobiliary Surgery, China-Japan Union Hospital of Jilin University, Changchun, China.
Eur Rev Med Pharmacol Sci. 2018 Sep;22(18):5899-5905. doi: 10.26355/eurrev_201809_15918.
Hepatocarcinoma is a great threat to global health. MicroRNA-23a was suggested to regulate growth and apoptosis in certain cell lines. Our study was focused on growth, proliferation, and apoptosis of hepatocarcinoma cell line MHCC97H under the influence of microRNA-23a, and explored the mechanism of pro-apoptosis microRNA-23a.
MicroRNA-23a and control microRNA (scramble miRNA, for short as miRNA) were synthesized with the routine protocol. Lipofection transfection was performed in hepatocarcinoma cell line MHCC97H. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, caspase-3 activity detection, and flow cytometry were performed to examine growth, proliferation, and apoptosis of hepatocarcinoma cell line MHCC97H, respectively. Kidney inhibitor of apoptosis protein (KIAP) and small interfere RNA (siRNA) was synthesized for inhibition of KIAP. KIAP plasmid was established for activation of KIAP. Western blot was performed to examine the protein expression of KIAP and caspase protein family after transfection of KIAP siRNA or KIAP plasmid.
Compared with miRNA transfection, microRNA-23a transfection significantly reduced the growth of MHCC97H cells, and decreased the expression of KIAP (p < 0.05). Enhanced translocation of phosphatidylserine and activation of caspase-3 were observed in microRNA-23a transfection cells. Moreover, inhibition of KIAP enhanced the pro-apoptosis effect of microRNA-23a, while activation of KIAP abrogated pro-apoptosis effect of microRNA-23a.
MicroRNA-23a inhibits growth and proliferation of MHCC97H cells, and induces apoptosis of MHCC97H cells via down-regulating KIAP. KIAP could be a potential therapeutic target for hepatocarcinoma treatment.
肝癌是全球健康的一大威胁。有研究表明 microRNA-23a 可调控某些细胞系的生长和凋亡。本研究旨在探讨 microRNA-23a 对肝癌细胞系 MHCC97H 的生长、增殖和凋亡的影响，并探索促凋亡 microRNA-23a 的作用机制。
采用常规方法合成 microRNA-23a 和对照 microRNA（简称 miRNA）。将脂质体转染法应用于肝癌细胞系 MHCC97H。采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）比色法、caspase-3 活性检测和流式细胞术分别检测肝癌细胞系 MHCC97H 的生长、增殖和凋亡。合成肾凋亡抑制蛋白（KIAP）和小干扰 RNA（siRNA）以抑制 KIAP，构建 KIAP 质粒以激活 KIAP。转染 KIAP siRNA 或 KIAP 质粒后，采用 Western blot 检测 KIAP 和 caspase 蛋白家族的蛋白表达。
与 miRNA 转染相比，microRNA-23a 转染明显抑制 MHCC97H 细胞的生长，并降低 KIAP 的表达（p<0.05）。microRNA-23a 转染细胞可见磷脂酰丝氨酸易位增强和 caspase-3 激活。此外，抑制 KIAP 增强了 microRNA-23a 的促凋亡作用，而激活 KIAP 则消除了 microRNA-23a 的促凋亡作用。
microRNA-23a 通过下调 KIAP 抑制 MHCC97H 细胞的生长和增殖，并诱导 MHCC97H 细胞凋亡。KIAP 可能成为肝癌治疗的潜在治疗靶点。
