The effects of miR-1243 on the proliferation and migration of hepatocellular carcinoma HepG2 cells through targeted regulation of hnRNPA2B1 expression / 中国肿瘤生物治疗杂志

@# [摘 要] 目的：探讨miR-1243通过靶向调控核不均一核糖核蛋白A2/B1（hnRNPA2B1）表达对肝癌HepG2细胞增殖、迁移的影响及其分子机制。方法：用qPCR和WB法检测40例肝癌组织及其癌旁组织（2019年1月至2021年8月在武汉市第三医院首义院区手术切除标本）和正常人肝细胞QSG-7701与肝癌细胞HepG2、Hep3b、HuH-7中miR-1243、hnRNPA2B1 mRNA水平及hnRNPA2B1、cyclin D1、MMP-2蛋白水平；双荧光素酶报告基因实验验证miR-1243和hnRNPA2B1的靶向关系。HepG2细胞分为对照组（不转染）、miR-NC组（转染miR-NC）、miR-1243 mimic组（转染miR-1243 mimic）、miR-1243 mimic+pcDNA3.1组（转染miR-1243 mimic和pcDNA3.1）、miR-1243 mimic+pc-hnRNPA2B1组（转染miR-1243 mimic和pc-hnRNPA2B1）后进行相应转染；WB法检测肝癌组织及细胞和转染后各组细胞的hnRNPA2B1、cyclin D1、MMP-2蛋白表达水平；CCK-8法检测转染后各组HepG2细胞的增殖能力；划痕愈合实验检测转染后各组HepG2细胞的迁移能力。结果：与癌旁组织或正常人肝细胞QSG-7701相比，肝癌组织和肝癌细胞中miR-1243呈低表达、hnRNPA2B1 mRNA及其蛋白呈高表达（均P<0.05）。双荧光素酶报告基因实验结果证实miR-1243与hnRNPA2B1间存在靶向关系，且miR-1243通过靶向hnRNPA2B1负调控其表达。转染miR-1243 mimic后HepG2细胞中hnRNPA2B1蛋白表达、细胞增殖能力、划痕愈合率及cyclin D1、MMP-2蛋白表达均显著降低（均P<0.05）；而同时过表达hnRNPA2B1和miR-1243可逆转过表达miR-1243对HepG2细胞增殖、迁移的抑制作用。结论：miR-1243通过靶向hnRNPA2B1表达调控肝癌HepG2细胞的增殖和迁移。

miR-502-3p regulates the proliferation and apoptosis of colorectal cancer stem cells by targeting GTPBP2 gene / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨miR-502-3p通过靶向GTP结合蛋白2（GTPBP2）调控结直肠癌干细胞（CCSC）增殖、细胞周期和凋亡的分子机制。方法：利用免疫磁珠分选技术从结直肠癌细胞HCT116中分选CCSC（CD133+CD44+双阳性细胞和CD133-CD44-双阴性细胞），用qPCR法检测细胞中miR-502-3p表达水平。利用脂质体转染法分别将miR-NC、miR-502-3p、si-miR-NC、si-miR-502-3p、miR-502-3p+vector和miR-502-3p+GTPBP2转染至CD133+CD44+细胞中，记作miR-NC、miR-502-3p、si-miR-NC、si-miR-502-3p、miR-502-3p+vector和miR-502-3p+GTPBP2组。用qPCR法检测细胞中miR-502-3p、GTPBP2 mRNA表达水平，MTT法、流式细胞术分别检测细胞增殖率、细胞周期和凋亡率，WB法检测细胞中Ki67、CDK1、Bcl2、BAX和GTPBP2蛋白的表达水平。双荧光素酶报告基因实验验证miR-502-3p与GTPBP2基因的靶向关系。结果：CD133+CD44+细胞中miR-502-3p表达水平显著低于CD133-CD44-细胞（P<0.01）。与miR-NC组比较，miR-502-3p组细胞增殖率、S期细胞比例显著降低（均P<0.01），凋亡率、G0/G1期细胞比例显著升高（均P<0.01），细胞中Ki67、CDK1、Bcl2蛋白表达均显著下调（均P<0.01）、BAX蛋白表达显著上调（P<0.01）。miR-502-3p靶向调控GTPBP2的表达，过表达GTPBP2可逆转上调miR-502-3p对CCSC增殖、周期和凋亡的作用。结论：上调miR-502-3p表达抑制CCSC增殖和阻滞细胞周期、诱导凋亡，其作用机制可能与过表达GTPBP2有关。

lncRNA ZFAS1 promotes the proliferation, invasion, and migration of liver cancer HepG2 cells through the miR-588/HMGA2 axis / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of long non-coding RNA zinc finger antisense 1 (lncRNA ZFAS1) on the proliferation, invasion, and migration of liver cancer cells by regulating the miR-588/high mobility group AT-hook protein 2 (HMGA2) axis. Methods: Real-time fluorescent quantitative PCR (qRT-PCR) and western blot were performed to measure the expression levels of ZFAS1, miR-588 and HMGA2 in 80 pairs of liver cancer tissues and corresponding para-cancerous issues (the tissue sample were collected from Shouyi Campus, Wuhan Third Hospital during Jan. 2018 and Dec. 2019), human normal liver cell line (LO2) and liver cancer cell lines (HepG2, Huh7, HCCLM3). The survival of patients was analyzed using the Kaplan-Meier survival curve. HepG2 cells were divided into blank group, si-NC group, si-ZFAS1 group, si-ZFAS1+inhibitor NC group, and si-ZFAS1+miR-588 inhibitor group. qPCR was performed to measure the expression of ZFAS1 and miR-588 in HepG2 cells of each group, and Western blot was performed to measure the expression of HMGA2 protein in the cells. CCK-8 method, Transwell, and scratch test were performed to measure the proliferation, invasion, and migration of HepG2 cells. Dual-luciferase reporter gene experiment was performed to verify the targeting relationship between ZFAS1 and miR-588 as well as between miR-588 and HMGA2. A HepG2 cell transplanted tumor model was established in nude mice to examine the effect of silencing ZFAS1 or/and miR-588 on the growth of transplanted tumors. Results: ZFAS1 and HMGA2 were highly expressed while miR-588 was lowly expressed in liver cancer tissues and liver cancer cells (all P<0.05). The 2-year survival rate of patients with low ZFAS1 expression was higher than that in the high expression group (P<0.05). Compared with the blank group, the relative expression of ZFAS1 and HMGA2 protein in the si-ZFAS1 group was significantly reduced, and the relative expression of miR-588 was significantly increased (all P<0.05); compared with the si-ZFAS1 group, the relative expression of ZFAS1 in the si-ZFAS1+miR-588 inhibitor group did not change significantly (P>0.05), however, the relative expression of HMGA2 protein was significantly increased, and the relative expression of miR-588 was significantly reduced (P<0.05). Silencing ZFAS1 was able to inhibit the proliferation, invasion, and migration of HepG2 cells and inhibit the growth of transplanted tumors in nude mice (all P<0.05). ZFAS1 targeted and down-regulated the expression of miR-588, while miR-588 targeted and down-regulated the expression of HMGA2. Simultaneous inhibition of miR-588 expression could reverse the inhibitory effects of silencing ZFAS1 on the proliferation, invasion, and migration of HepG2 cells and the growth of transplanted tumors in nude mice (all P<0.05). Conclusion: Silencing ZFAS1 may down-regulate the expression of HMGA2 by promoting the expression of miR-588, thereby inhibiting the proliferation, invasion, and migration of liver cancer HepG2 cells.