Expression Of KRT17 In The Keratinocytes Of Condyloma Acuminatum And Its Role In Proliferation And Apoptosis

Background: Condyloma acuminata is a genital wart caused by HPV infection.A large number of studies have suggested the keratinocytes in condyloma acuminatum proliferated abnormally,but the mechanism of HPV-induced keratinocyte dysplasia is still unclear.Studies have found that keratinocytes in condyloma acuminatum had a high level of expression of KRT17,and other studies have shown that the expression of KRT17 in cervical cancer cells induced by high-risk type HPV is related to cell proliferation.Because keratinocyte abnormal hyperplasia is the basis of the formation of wart,deeply exploring the role of KRT17 in keratinocytes of condyloma acuminatum is conducive to understand the mechanism of condyloma acuminatum,thus beneficial to prevent and cure condyloma acuminatum.Objectives: To investigate the expression of KRT17 and its role in proliferation and apoptosis in HPV infected keratinocytes and non HPV infected keratinocytes of condyloma acuminatum.Methods:(1)Datasets of human specimens,including condyloma acuminatum and foreskin,obtained from GEO datasets were used to analyze the mRNA expression of KRT17,and further confirmed the results of datasets by samples of condyloma acuminatum with Real-time quantitative PCR;(2)Immunohistochemistry was used to detect the expression of KRT17 and proliferation index(Ki67)in condyloma acuminatum and western blot was used to detect the expression of KRT17,then analyze its relationship with proliferation and with parameter of patients with condyloma acuminatum;(3)A cell model of condyloma acuminatum,HPV11-E7 HaCaT was constructed by lentivirus with HPV11-E7 and the overexpression of HPV11-E7 can induce the high expression of KRT17.siRNA was used to knock down the expression of KRT17 in HPV11-E7 HaCaT,whose proliferation and apoptosis were detected;(4)Immunofluorescence staining of phalloidin was performed to showed the change of cytoskeletal organization when KRT17 was knocked down in HPV11-E7 HaCaT and flow cytometry was used to investigate the change of cell cycle;(5)The medium from cell model with KRT17 siRNA or control siRNA was collected,making it conditional medium by filtering it,then used fresh feed HaCaT,and the change in proliferation and apoptosis of HaCaT were detected.Results:(1)mRNA microchips showed that the expression of KRT17 in condyloma acuminatum was higher than foreskin and specimens of condyloma acuminatum confirmed the results of microchips.(2)Higher expression of KRT17 and Ki67 were found in condyloma acuminatum and we discovered that a positive correlation between KRT17 expression and proliferation of keratinocytes in condyloma acuminatum.Besides,we explored the relationship between KRT17 expression and course of disease: in lesion with short period,KRT17 expression was higher,but no significant correlation between KRT17 expression and age,gender or location were observed.(3)After KRT17 was knocked down,the cell model of condyloma acuminatum showed a decrease in proliferation and an increase in apoptosis.(4)Cytoskeleton protein F-actin did not change,but the S phase cell cycle was significantly prolonged when KRT17 was knocked down.(5)After treatment with conditional medium from HPV11-E7 HaCaT with KRT17 knockdown,HaCaT showed a decrease in proliferation and an increase in apoptosis,indicating that KRT17 might affect cell by regulating the secretion of cytokines.Conclusion: In keratinocytes infected with HPV,KRT17 regulated the proliferation and apoptosis of cell by regulating the cell cycle.Meanwhile,by regulating the secretory cytokines,KRT17 can promote the proliferation of normal keratinocytes and inhibit apoptosis,thus warts were formed.