HOXA10 knockdown inhibits proliferation, induces cell cycle arrest and apoptosis in hepatocellular carcinoma cells through HDAC1
HOXA10 knockdown inhibits proliferation, induces cell cycle arrest and apoptosis in hepatocellular carcinoma cells through HDAC1
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HOXA10敲低通过HDAC1抑制肝细胞癌细胞增殖、诱导细胞周期停滞和凋亡
DOI：
10.2147/cmar.s199239
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发表时间：
2019-01-01
影响因子：
3.3
通讯作者：
Zang, Guo-Qing
中科院分区：
文献类型：
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作者：
Zhang, Yi;Chen, Jie;Zang, Guo-Qing
背景同源盒A10(Homeobox A10，HOXA10)与多种人类肿瘤的发生发展密切相关。然而，HOXA10在肝细胞癌中的确切生物学功能尚未明确。方法采用实时定量聚合酶链式反应(qRT-PCR)检测肝细胞癌组织和细胞株中HOXA10基因的表达，Western印迹法检测组蛋白脱乙酰酶(HDAC)、组蛋白脱乙酰酶(HDAC)、细胞周期蛋白D1、增殖细胞核抗原(PCNA)、Survivin和P53乙酰化水平。用细胞计数试剂盒(CCK-8)检测细胞增殖，用流式细胞仪分析细胞周期。此外，利用裸鼠移植瘤模型监测了肝癌细胞在体内的生长情况。用荧光素酶报告素法和染色质免疫沉淀法分别检测HDAC1启动子在肝癌细胞系中的活性和结合情况。结果我们发现HOXA10在肝细胞癌组织中的表达高于癌旁正常肝组织。RNA干扰介导的HOXA10基因敲除在体内外均能抑制肝癌细胞的增殖。HOXA10基因敲除后，细胞周期停滞于G0/G1期，细胞发生凋亡，细胞周期蛋白Cyclin D1、增殖细胞核抗原和Survivin表达降低。值得注意的是，HOXA10基因敲除增强了P53的乙酰化(Lys382)，这对P53的激活至关重要。同样，HOXA10基因敲除抑制了HDAC1的转录，HDAC1是一种潜在的P53脱乙酰酶。与此相一致，HDAC1下调HOXA10过表达对细胞增殖、细胞周期进程、细胞凋亡和p53乙酰化的影响，表明HDAC1在HOXA10功能调节中的作用。结论HOXA10基因敲除可能通过调控HDAC1的转录抑制肝癌细胞的增殖，诱导细胞周期停滞和凋亡。
Background Homeobox A10 (HOXA10) has been implicated in the development and progression of various human cancers. However, the precise biological functions of HOXA10 in hepatocellular carcinoma (HCC) have not been defined. Methods In this study, we examined mRNA expression by quantitative real-time PCR (qRT-PCR) of HOXA10 as well as histone deacetylase (HDAC) and protein levels by Western blot of HOXA10, HDAC1, Cyclin D1, proliferating cell nuclear antigen (PCNA), Survivin and p53 acetylation in HCC tissues and cell lines. We also assessed cell proliferation using Cell Counting Kit-8 (CCK-8) and analyzed cell cycle by flow cytometry. Furthermore, tumor growth of HCC cells in vivo was monitored using the nude mouse xenograft model. Finally, HDAC1 promoter activity and binding in HCC cell lines were detected by luciferase reporter assay and chromatin immunoprecipitation (ChIP), respectively. Results We uncovered the elevated expression of HOXA10 in HCC tissues compared to adjacent normal liver tissues. RNA interference-mediated knockdown of HOXA10 inhibited HCC cell proliferation both in vitro and in vivo. HOXA10 knockdown also induced cell cycle arrest at G0/G1 phase and apoptosis, which were accompanied with the reduced expression of Cyclin D1, PCNA and Survivin. Notably, HOXA10 knockdown enhanced p53 acetylation (Lys382), which is crucial to the activation of p53. Likewise, HOXA10 knockdown suppressed the transcription of HDAC1, a potential deacetylase for p53. In line with these observations, HDAC1 downregulation abrogated the effects of HOXA10 overexpression on proliferation, cell cycle progression, apoptosis and p53 acetylation, indicating the role of HDAC1 in mediating HOXA10 functions. Conclusion Our results demonstrate that HOXA10 knockdown inhibits proliferation, induces cell cycle arrest and apoptosis in HCC cells by regulating HDAC1 transcription.