Functional And Ralated Mechanistic Studies Of NUP93 In Maligant Progress In Cervical Cancer

Objective:To analyze the expression of NUP93 in cervical cancer tissues and normal cervical tissues,exploring the malignant biological behaviors such as cellular proliferation,metastasis and invasion ability after silencing NUP93 in cervical cancer cells and related mechanism.Methods:(1)We analyzed the expression of NUP93 in cervical cancer tissues and normal cervical tissues using immunohistochemical staining,discussing the relationship between NUP93 expression and the clinical pathological parameters.We examined the the expression of Nup93 in cervical cancer cell lines at the mRNA level by qPCR and at the protein level by Western blotting.(2)We detected cellular proliferation ability after silencing NUP93 in cervical cancer cell lines(SiHa,C33a)using the CCK-8 assay and Colony formation assay.(3)We detected the capabilities of migration and invasion after silencing NUP93 in cervical cancer cells(SiHa,C33a)using the wound healing test and Transwell assay.(4)We detected the cell cytoskeleton after silencing NUP93 in cervical cancer cells(SiHa,C33a)using the F-actin staining.(5)We analyzed the tumor formation in vivo after silencing NUP93 in cervical cancer cells(SiHa,C33a)by subcutaneous transplantation of the tumor.(6)We detected the expression of the related proteins of IL-6/STAT3 pathway in cervical cancer cells(SiHa,C33a)using Western blotting.We analyzed the expression of the related proteins of IL-6/STAT3 pathway after silencing NUP93 in cervical cancer cells(SiHa,C33a).Results(1)Nup93 expression was higher in cervical cancer tissues than that in cancer adjacent cervical tissues(P<0.05).Nup93 was mainly localized on the nuclear membrane.The differently expression of Nup93 between squamous cell carcinoma and adenosquamous carcinoma was not significant(P>0.05).The immunostaining of Nup93 was higher in cancer tissues in advanced stages(stage III/IV)than those in the early stages(stage I/II)(P<0.05).The immunostaining of Nup93 was higher in cancer tissues in advanced grades than those in the early grades(grades 2-3 vs 1,P<0.05).However,the relation of Nup93 expression with age was not significant(Table I;P>0.05).(2)The down-regulation of Nup93 in SiHa and C33 a cell lines resulted in a decrease in cell proliferation compared with LV-3-NC cell lines by the CCK-8 assay and the colony formation assay.(3)The migration and invasion ability of LV3-Nup93 infected SiHa and C33 a cells was suppressed compared with LV3-NC infected cells by the the wound healing test and Transwell assay.(4)In LV3-NC infected SiHa and C33 a cells,F-actin staining was mainly localized in the cellular outgrowth and projections.On the contrary,in LV3-Nup93 infected SiHa and C33 a cells,F-actin staining was homogenic across the cytoplasm,and the formation of membrane ruffles and lamellipodia was prevented.(5)The mean weight and volume of tumors in LV3-Nup93 infected group was lower than that of the LV3-NC infected group(P<0.05).(6)The silencing of Nup93 in SiHa and C33 A cells caused a reduction in protein levels of P-STAT3 but it can not influence the level of IL-6R.IL-6 could restore the expression of Nup93 and P-STAT3 in the LV3-Nup93 infected SiHa and C33 A cells.The findings suggested that IL-6 could regulate the expression of Nup93 and P-STAT3.And Nup93 interacted with IL-6/STAT3 pathway.ConclusionNUP93 is a tumor promotor which can regulate cell cytoskeleton,cellular proliferation,migration and invasion ability by IL-6/STAT3 pathway in cervical cancer.