Regulation Of ATP5B On Biological Behavior Of Cervical Carcinoma HeLa Cells

ObjectiveATP5B gene encodes beta subunit of mitochondrial ATP synthase. Abnormal expression of β subunit would affect the function of ATP synthase. Research shows that more and more diseases are associated with the abnormal expression of ATP synthase. ATP synthase has also been demonstrated and suggested as a good molecular target for drugs in the treatment of cancer. However, the expression level of ATP5B in cervical carcinoma has not been reported until noW, and its biological function is not well known. This paper is to study the expression level and biological functions of ATP5B in cervical carcinoma HeLa cells. In addition, its potential mechanisms were also involved.MethodsReal-time PCR was performed to detect the expression level of ATP5B in six pairs of cervical carcinoma tissue samples and matched normal cervical tissue samples. Constructing the recombinant vector pRNAT-U6.2/ATP5B-shRNA, which was transfected into HeLa via lipofectamine. Western Blotting detected the interference of pRNAT-U6.2/ATP5B-shRNA on ATP5B. MTT assay, clone information assay and growth curve, monolayer wound assay, transwell invasion assay, aggregation assay were used to analyze the silencing of ATP5B on cell viability, proliferation capacity, migration, invasion and adhesion. The effect of ATP5B silencing on the sensitivity of HeLa to paclitaxel was detected by MTT assay. The cell cycle and apoptosis were detected by FACS after knockdown of ATP5B.ResultsATP5B was up-regulated in cervical carcinoma tissue, compared to normal cervical tissue. The recombinant vector pRNAT-U6.2/ATP5B-shRNA was successfully constructed and the interfering efficiency of pRNAT-U6.2/ATP5B-shRNA on ATP5B was about70.1%. The down-regulation of ATP5B inhibited HeLa cell viability, proliferation capacity, migration and invasion, but increased the cell adhesion. Furthermore, knockdown of ATP5B significantly sensitized HeLa cells for paclitaxel-induced cytotoxicity. The FACS assay showed that the knockdown of ATP5B did not arrest the cell cycle, but induce cell apoptosis.ConclusionATP5B is up-regulated in cervical carcinoma tissue. The down-regulation of ATP5B inhibited cell viability, proliferation capacity, migration and invasion and increased the cell adhesion in cervical carcinoma cells. ATP5B gene silencing increased the sensitivity of HeLa to paclitaxel. The knockdown of ATP5B is shown to be able to significantly prevent cell proliferation and invasion through the promotion of cell apoptosis, rather than cell cycle arrest. ATP5B has played important and complex roles in the development of cervical carcinoma, which provided a new clue to the research of cancer initiation, progression, diagnosis and therapy.