Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados (CINVESTAV) AP, 14740, Mexico.
Laboratorio de Biomedicina Molecular, Unidad Académica de Ciencias Químico Biológicas, Universidad Autónoma de Guerrero, Chilpancingo, Mexico.
Virus Res. 2018 Mar 2;247:94-101. doi: 10.1016/j.virusres.2018.02.009. Epub 2018 Feb 13.
The HPV-16 E6/E7 bicistronic immature transcript produces 4 mature RNAs: the unspliced HPV-16 E6/E7 product and 3 alternatively spliced mRNAs. The 3 spliced mRNAs encode short forms of the E6 oncoprotein, namely E6I, E6II and E6^E7. In this study we showed that transfection of C-33A cells with monocistronic constructs of these cDNAs fused to GFP, produced different effects on apoptosis, after the treatment with cisplatin. Transfection of C-33A cells with the full-length E6-GFP oncoprotein resulted in a 50% decrease in cell death, while the transfection with the E6I-GFP construct showed only a 25% of diminution of cell death, compared to the control cells. Transfection with the E6^E7-GFP or E7-GFP construct had no effect on the number of the apoptotic cells, compared with control cells. Conversely, transfection with the E6II construct resulted in higher cell death than the control cells. Taken together, these results suggested that E6I or E6II, the short forms of HPV-16 E6, displayed opposite effects on cisplatin-induced apoptosis, when transfected in C-33A cells.
HPV-16 E6/E7 双顺反子不成熟转录物产生 4 种成熟 RNA：未剪接的 HPV-16 E6/E7 产物和 3 种选择性剪接的 mRNA。这 3 种剪接的 mRNA 编码 E6 癌蛋白的短形式，即 E6I、E6II 和 E6^E7。在这项研究中，我们表明，用这些 cDNA 的单顺反子构建体转染 C-33A 细胞，并融合 GFP，在用顺铂处理后，对细胞凋亡产生不同的影响。全长 E6-GFP 致癌蛋白的转染导致细胞死亡减少 50%，而 E6I-GFP 构建体的转染仅导致细胞死亡减少 25%，与对照细胞相比。与对照细胞相比，用 E6^E7-GFP 或 E7-GFP 构建体转染对凋亡细胞的数量没有影响。相反，E6II 构建体的转染导致细胞死亡高于对照细胞。总之，这些结果表明，HPV-16 E6 的短形式 E6I 或 E6II，在转染 C-33A 细胞时，对顺铂诱导的细胞凋亡表现出相反的影响。
