Department of Clinical Biochemistry, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
Cellular and Molecular Research Center, Urmia University of Medical Sciences, Urmia, Iran.
Med Oncol. 2022 Aug 16;39(11):165. doi: 10.1007/s12032-022-01745-5.
Hepatocellular carcinoma (HCC), the most prevalent kind of liver cancer, remains one of the world's main causes of death. The alpha7 nicotinic acetylcholine receptor (α7nAchR) has been recognized to be overexpressed in malignancies and chemoresistance. Since little is known about the role of α7nAchR expression in drug-resistant cells, this study was designed to investigate the effect of α7nAchR suppression in combination with Sorafenib (SOR) on SOR-resistant HCC cells. First, SOR-resistant HCC cells were generated. To suppress the expression of α7nAchR, cells were treated with SOR following siRNA transfection. qRT-PCR was used to examine the expression of α7nAchR and apoptotic genes by evaluating the IC50 of SOR and the combination of α7nAchR siRNA and SOR on the survival of resistant cells. Moreover, apoptosis, autophagy, and cell cycle analysis for resistant HCC cells were performed using flow cytometry. Cell migration and colony formation assays were also used for further confirmation. Our results suggest that inhibiting α7nAchR can lead resistant HCC cells to become sensitive. Furthermore, when siRNA and SOR were treated together, HCC-resistant cells showed a considerable reduction in α7nAchR mRNA gene expression. In addition, when α7nAchR was downregulated in combination with SOR, migration and colony formation were inhibited. Apoptosis was triggered by modulating the expression of apoptotic target genes, and cell cycle arrest was observed in the G2-M and subG1 phases. Overexpression of α7nAchR in SOR-resistant HCC cells suggests that it might be a therapeutic target for HCC cell resistance therapy.
肝细胞癌（HCC）是最常见的肝癌类型，仍是全球主要死亡原因之一。α7 烟碱型乙酰胆碱受体（α7nAchR）在恶性肿瘤和化疗耐药中被认为过度表达。由于对 α7nAchR 表达在耐药细胞中的作用知之甚少，因此本研究旨在研究抑制α7nAchR 与索拉非尼（SOR）联合用药对 SOR 耐药 HCC 细胞的影响。首先，生成 SOR 耐药 HCC 细胞。为了抑制 α7nAchR 的表达，用 SOR 处理细胞，然后用 siRNA 转染。通过评估 SOR 和 α7nAchR siRNA 与 SOR 联合对耐药细胞存活的 IC50，使用 qRT-PCR 检测 α7nAchR 和凋亡基因的表达。此外，使用流式细胞术对耐药 HCC 细胞进行凋亡、自噬和细胞周期分析。还进行了细胞迁移和集落形成测定以进一步确认。我们的结果表明，抑制 α7nAchR 可使耐药 HCC 细胞变得敏感。此外，当用 siRNA 和 SOR 联合处理时，HCC 耐药细胞中 α7nAchR mRNA 基因表达显著降低。此外，当与 SOR 一起下调 α7nAchR 时，迁移和集落形成受到抑制。通过调节凋亡靶基因的表达触发凋亡，并观察到细胞周期停滞在 G2-M 和 subG1 期。SOR 耐药 HCC 细胞中 α7nAchR 的过表达表明，它可能是 HCC 细胞耐药治疗的一个治疗靶点。
