Department of Burn and Plastic Surgery, Cangzhou Central Hospital, Cangzhou City, Hebei Province, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(3):965-971. doi: 10.26355/eurrev_201902_16983.
Connexin 43 (Cx43), a vital gap junction protein is reported to be involved in melanoma progression. The aim of the study is to investigate the regulatory role of Cx43 in melanoma.
Western blot assay was used to detect the protein expression of Cx43 in melanoma cells and the human epidermal melanocytes (HEMn). MTT cell proliferation and cell colony formation assays were used to assess cell proliferation. Bioinformatics prediction, luciferase reporter assay, Western blot and qRT-PCR assays were applied to demonstrate that Cx43 was a direct target of miR-106a in melanoma cells.
Connexin 43 (Cx43) was lower expressed in melanoma cells compared with human epidermal melanocytes (HEMn). Cx43 overexpression significantly inhibited melanoma cell proliferation and colony formation ability in vitro. However, knockdown of Cx43 had opposite effects on cell proliferation and colony formation. Bioinformatics prediction and luciferase reporter assays demonstrated that miR-106a targeted the 3' untranslated region (3'UTR) of Cx43 and regulated its mRNA and protein expression levels in melanoma cells. MiR-106a was upregulated in melanoma cells, and its overexpression attenuated the effects caused by upregulating Cx43 expression.
Thus, our results indicated that Cx43 was downregulated in melanoma cells and may be a potential target of melanoma treatment.
连接蛋白 43(Cx43)是一种重要的间隙连接蛋白,据报道其参与了黑色素瘤的进展。本研究旨在探讨 Cx43 在黑色素瘤中的调控作用。
Western blot 检测黑色素瘤细胞和人表皮黑素细胞(HEMn)中 Cx43 的蛋白表达。MTT 细胞增殖和细胞集落形成实验用于评估细胞增殖。生物信息学预测、荧光素酶报告实验、Western blot 和 qRT-PCR 实验用于证明 Cx43 是黑色素瘤细胞中 miR-106a 的直接靶标。
与 HEMn 相比,黑色素瘤细胞中 Cx43 的表达水平较低。Cx43 过表达显著抑制黑色素瘤细胞的体外增殖和集落形成能力。然而,Cx43 的敲低对细胞增殖和集落形成有相反的影响。生物信息学预测和荧光素酶报告实验表明,miR-106a 靶向 Cx43 的 3'非翻译区(3'UTR),并调节其在黑色素瘤细胞中的 mRNA 和蛋白表达水平。miR-106a 在黑色素瘤细胞中上调,其过表达减弱了上调 Cx43 表达引起的作用。
因此,我们的结果表明 Cx43 在黑色素瘤细胞中下调,可能是黑色素瘤治疗的潜在靶点。