Mosweu Mpho, Motadi Lesetja, Moela Pontsho
University of Pretoria, Faculty of Natural and Agricultural Sciences, Department of Biochemistry, Hatfield, Pretoria 0002, South Africa.
University of Johannesburg, Faculty of Science, Department of Biochemistry, Auckland Park Campus, Auckland Park, Johannesburg 2050, South Africa.
Cancer Manag Res. 2020 Oct 29;12:10725-10734. doi: 10.2147/CMAR.S261576. eCollection 2020.
is one of the genes identified as a proliferative gene that plays a role in cancer development, On the other hand both RBBP6 and telomerase activity have been shown to be increase in various cancers. E6 protein of HPV and RBBP6 is known to enhance the progression of cancer cells by interacting with p53 and presenting it to be ubiquitinated by the proteasome thereby promoting cell proliferation and preventing apoptosis. Studies also show that HPV E6 protein can increase telomerase activity by activating the expression of human telomerase reverse transcriptase (hTERT), thus enabling the immortalization of the cells. With RBBP6 and hTERT showing a similar profile in cancer cells, we seek to investigate any possible effect of RBBP6 on telomerase activity.
Using real-time qPCR and TRAPeze RT Telomerase detection kit (Merc) respectively. We used cervical cancer cell lines in which CaSki cell showed the reduction of hTERT expression and reduction in telomerase activity significantly in RBBP6-knockdown cells. While no significant changes were observed in HeLa cells. Real-time growth assay revealed a significant drop in cell growth in co-silenced RBBP6 and hTERT cells substantiating our observation that RBBP6 might be playing a role in cell proliferation.
Taken all together, the observed effect of RBBP6 gene silencing on telomerase activity in cervical cancer is cell line dependent.
是被鉴定为在癌症发展中起作用的增殖基因之一。另一方面,RBBP6和端粒酶活性在各种癌症中均显示增加。已知人乳头瘤病毒(HPV)的E6蛋白和RBBP6通过与p53相互作用并使其被蛋白酶体泛素化,从而促进癌细胞增殖并防止细胞凋亡,进而增强癌细胞的进展。研究还表明,HPV E6蛋白可通过激活人端粒酶逆转录酶(hTERT)的表达来增加端粒酶活性,从而使细胞永生化。鉴于RBBP6和hTERT在癌细胞中表现出相似的特征,我们试图研究RBBP6对端粒酶活性的任何可能影响。
分别使用实时定量PCR和TRAPeze RT端粒酶检测试剂盒(Merc)。我们使用了宫颈癌细胞系,其中CaSki细胞在RBBP6基因敲低的细胞中hTERT表达降低且端粒酶活性显著降低。而HeLa细胞未观察到显著变化。实时生长分析显示,RBBP6和hTERT共同沉默的细胞中细胞生长显著下降,证实了我们关于RBBP6可能在细胞增殖中起作用的观察。
综上所述,RBBP6基因沉默对宫颈癌中端粒酶活性的观察到的影响是细胞系依赖性的。