Department of Hepatobiliary Surgery, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
Eur Rev Med Pharmacol Sci. 2018 Oct;22(20):6784-6792. doi: 10.26355/eurrev_201810_16145.
We investigate the mechanism of HOXB-AS3 in promoting the development of hepatocellular carcinoma.
The expression of HOXB-AS3 in tumor tissues and adjacent tissues of hepatocellular carcinoma was detected by quantitative real time-polymerase chain reaction (qRT-PCR), and the relationship between the expression of HOXB-AS3 and tumor tissues was analyzed. The effects of HOXB-AS3 and p53 on cell proliferation, cell cycle and apoptosis were detected by plate cloning experiment and flow cytometry. The binding relationship between HOXB-AS3 and DNMT1 and the regulation mechanism of DNMT1 on p53 were tested by RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) experiments, respectively. Western blot was used to detect the expression of p53 after knockdown of HOXB-AS3. The torsion experiment was performed to assess whether HOXB-AS3 regulated the proliferation and apoptosis of hepatoma cells by inhibiting p53 expression.
The results of qRT-PCR showed that the expression of HOXB-AS3 was significantly higher in cancerous tissues of patients with hepatocellular carcinoma than in adjacent tissues. The expression of HOXB-AS3 in patients in stage III and IV was significantly higher than that in stage I and II. Inhibition of HOXB-AS3 expression in liver cancer cells including Hep3B and LM3 could promote cell proliferation, inhibit cell apoptosis and induce cell cycle arrest in the G0/G1 phase. The results of RIP and ChIP experiments showed that HOXB-AS3 inhibited the expression of p53 by binding to DNMT1, and overexpression of p53 in Hep3B cells could partially reverse the changes in cell proliferation and apoptosis induced by HOXB-AS3.
Highly expressed HOXB-AS3 was confirmed to promote the proliferation of hepatocellular carcinoma cells and inhibit apoptosis, and the mechanism was related to the regulation role of HOXB-AS3 in p53 expression by binding to DNMT1.
研究 HOXB-AS3 在促进肝细胞癌发展中的作用机制。
采用实时荧光定量聚合酶链反应（qRT-PCR）检测肝癌组织及癌旁组织中 HOXB-AS3 的表达情况，并分析其与肿瘤组织的关系。采用平板克隆实验和流式细胞术检测 HOXB-AS3 和 p53 对细胞增殖、细胞周期和细胞凋亡的影响。分别采用 RNA 免疫沉淀（RIP）和染色质免疫沉淀（ChIP）实验检测 HOXB-AS3 与 DNMT1 的结合关系以及 DNMT1 对 p53 的调控机制。采用 Western blot 检测 HOXB-AS3 敲低后 p53 的表达情况。采用扭转实验评估 HOXB-AS3 是否通过抑制 p53 表达来调节肝癌细胞的增殖和凋亡。
qRT-PCR 结果显示，肝细胞癌患者癌组织中 HOXB-AS3 的表达明显高于癌旁组织。Ⅲ期和Ⅳ期患者 HOXB-AS3 的表达明显高于Ⅰ期和Ⅱ期患者。抑制肝癌细胞 Hep3B 和 LM3 中 HOXB-AS3 的表达可促进细胞增殖，抑制细胞凋亡，并诱导细胞周期停滞在 G0/G1 期。RIP 和 ChIP 实验结果表明，HOXB-AS3 通过与 DNMT1 结合抑制 p53 的表达，过表达 Hep3B 细胞中的 p53 可部分逆转 HOXB-AS3 诱导的细胞增殖和凋亡变化。
高表达的 HOXB-AS3 被证实可促进肝细胞癌细胞的增殖并抑制凋亡，其机制与 HOXB-AS3 通过与 DNMT1 结合调节 p53 表达有关。
