Kazrin F Promotes Malignancy Of Cervical Cancer Cell

ObjectiveKazrin protein was first found in 2004 by Karen R.Groot and his colleagues.They had found four isomers of kazrin A,kazrin B,kazrin C,kazrin D(amino acid sequences of kazrin C and kazrin D are same),and prove it's associated with cell connections.Then,there are several research on kazrin,in which it is associated with the mouse and Xenopus embryonic development.In 2009,Rachida Nachat discovered another isomer kazrin E,and proved its link with microtubule,related to epidermal differentiation and desmosome formation.The same year,our laboratory found another novel isoform kazrin F,has anti-apoptotic effects.This topic plans to investigate the effect of kazrin F on malignant behavior of cervical cancer cells based on the previous work.Cervical cancer is a type of women cancer with higher morbidity and mortality around the world,especially in developing countries.Through this study we try to elucidate whether kazrin F may service as an novel biomarker or target in diagnosis and treatment of cervical cancer.MethodWe first apply qRT-PCR to detect the mRNA expression level of kazrin F in cervical cancer tissues,preliminary forecast the possible role of kazrin F in tumorigenesis.Then we respectively over-express or knockdown kazrin F in cervical carcinoma cell HeLa,MTT assay,plate clone formation assay,transwell migration and invasion assay were used to detect the influence of kazrin F on cell viability,growth,proliferation ability,migration and invasion ability in cervical cancer cells.Afterwards,in order to elucidate the mechanism of kazrin F cell growth and proliferation ability,flow cytometry was used to test the effect of kazrin F on cell cycle and apoptosis of cervical cancer cells.Western blot was used to detect effect of kazrin F on epithelial mesenchymal transition of cervical cancer cells,to clarify the mechanism of kazrin F effect on cell migration and invasion.Finally,we tested the effect of kazrin F on cervical carcinoma cell autophagic capacity by fluorescent microscope and Western blot experiments.ResultqRT-PCR detection shows high expression of kazrin F in cervical cancer tissues compared with normal cervical tissues.After verification effectiveness of kazrin F overexpression and knockdown plasmid,MTT detected that kazrin F can promote cervical cancer cell viability;plate clone formation assay demonstrated that kazrin F could promote cervical cancer cell growth and preliferation ability;transwell migration assay and invasion assay confirmed that kazrin F can promote the migration and invasion ability of cervical carcinoma cells.Flow cytometry detection demonstrated kazrin F can inhibit the apoptosis induced by paclitaxel in cervical cancer cells,promote the process of cell cycle.Western blot were used to detect epithelial markers and mesenchymal markers,found that kazrin F could promote epithelial mesenchymal transition of cervical cancer cells.Finally,fluorescence microscopic observation and western blot detection confirmed that kazrin F can improve exogenous and endogenous LC3-II levels in cervical cancer cells,promote cervical cancer cell autophagy.ConclusionThses results indicate that kazrin F function as oncogene in cervical cancer,which promotes cell gowth and migration/invasion by modulating the cell cycle,apoptosis,EMT and autophagy.It may have potential value for diagnosis and therapeutics as a new biomarker.