Effects of HPV16E6 transfection on the biological behavior of Eca109 and Eca9706 cells.

Abstract

The aim of the present study was to investigate the effect of HPV16E6 gene integration on the biological behavior of Eca109 and Eca9706 cells. This was evaluated through positive liposome transfection of HPV16E6 into Eca109 cells and Eca9706 cells. The transfection efficiency was evaluated by calculating the ratio of fluorescent cells to total cells. After stable screening, reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the target gene and HPV16E6mRNA inside the cells. The distribution of HPV16E6 in esophageal carcinoma cells was observed by immunofluorescence and western blot analysis. A CCK-8 assay was performed to detect cell proliferation. The migration rate was measured by a wound healing assay, and a Transwell Matrigel invasion assay was used to detect invasive ability. The results of RT-PCR, immunofluorescence and western blot analyses indicated that HPV16E6 gene was expressed in the target group. The proliferation rates and clone group numbers were significantly higher in HPV16E6-transfected cell groups compared with nonsense-transfected (negative control) cell groups. The wound healing and Transwell invasion assays indicated that the migration rate and invasive ability were also significantly higher in the HPV16E6-transfected cell groups compared with negative control groups. In conclusion, Eca109 and Eca9706 cell lines with integration of HPV16E6 were successfully established in the present study. It was demonstrated that HPV16E6 expression enhanced the proliferation and migration of esophageal cancer cells. HPV16E6 may serve a key function in the occurrence and development of esophageal cancer.