Mfn2 Promotes Proliferation,Migration And Invasion Of Cervical Cancer Cells Through IL-6/STAT3 Signaling Pathway

Cervical cancer is one of the most common malignancies in the female reproductive system.The common pathological types of cervical cancer are: cervical squamous cell carcinoma,cervical adenocarcinoma,cervical squamous cell carcinoma and undifferentiated carcinoma,which cervical squamous cell scale is the most common type.In developed countries,cervical squamous cell carcinoma is not only the highest incidence tumor of gynecological reproductive system tumors,but also is second lethality in gynecological tumors,second only to ovarian cancer mortality.So far,the mechanism of cervical cancer metastasis is still not entirely clear.So to find the impact of cervical cancer invasion and metastasis of molecular markers and its invasion and metastasis mechanism,is the current focus of cervical cancer research.The Mfn2 protein is a mitochondrial protein encoded by the Mfn2 gene on the short arm of human chromosome 1,it contains 19 exons that encode a protein containing 757 amino acids.Mfn2 is distributed in mitochondria of human tissues and tissues,including female genital tract,cervical cells and so on.It can regulate and stabilize the local microenvironment state by regulating mitochondrial fusion strength.According to different types of tumors and tumor pathology,Mfn2 plays a different role and can promote or inhibit the occurrence of malignant tumors.There has been no study on the expression and related effects of Mfn2 in cervical malignancies.Therefore,this study through the following three parts of Mfn2 in cervical cancer expression and its proliferation,migration and invasion of the role and its potential molecular mechanisms were discussed.Part I The detection of Mfn2 in cervical cancer and cervical cancer cells in the expressionObjective: To investigate the expression of Mfn2 in cervical cancer and cervical cancer cell lines and its correlation with clinicopathological parameters.Methods: The expression of Mfn2 in cervical cancer tissues and cell lines was detected by immunohistochemistry and Western blotting.The expression of Mfn2 in different cervical cancer cell lines was detected by Western blotting.Results: The expression of Mfn2 in cervical cancer was significantly higher than that in adjacent normal cervical tissues(P <0.05).There was no significant difference in the expression of Mfn2 between squamous cell carcinoma and adenosquamous carcinoma(P> 0.05).The expression of Mfn2 was not significantly related to age(P> 0.05;Table 1).The expression of Mfn2 was associated with grading of cervical cancer and was not correlated with stage(grades 2-3 vs 1,P <0.05;III / IV vs I / II,P> 0.05).Conclusion: The expression of Mfn2 in cervical adenocarcinoma was significantly increased.Highly expressed Mfn2 is associated with grading of cervical cancer,independent of age and pathological stage,and Mfn2 is highly expressed in clinical early metastasisPART II investigate the effect and mechanism of Mfn2 on the proliferation,migration and invasion of cervical cancer cell linesObjective: To investigate the effect of Mfn2 on the proliferation,migration and invasion of cervical cancer cell lines(Si Ha and C33a)in cervical cancer.Methods: LV3-Mfn2 was transfected into cervical cancer cell line by lentivirus transfection technique.The proliferation ability of cervical cancer cells after silencing Mfn2 was detected by CCK-8.The migration ability of cervical cancer cells after silencing Mfn2 was detected by scratch test The changes of invasive ability of cervical cancer cells after silencing Mfn2 were detected by Transwell assay.Results: After transfection of LV3-Mfn2 lentivirus,the transfection efficiency of Si Ha and C33 a was observed by fluorescence microscopy,which was more than 90%.After transfection of LV3-Mfn2 lentivirus,CCK-8 and clone formation experiments showed that the proliferation of cervical cancer cell lines.We used CCK-8 kit to detect cell proliferation,the results shown in Figure 4,LV3-Mfn2 cells compared with LV-NC cells showed decreased proliferation of cells.Conclusion: The expression of Mfn2 in cervical adenocarcinoma was significantly increased.Highly expressed Mfn2 is associated with grading of cervical cancer,independent of age and pathologic stage,and Mfn2 expression is associated with early clinical metastasis.PART III Investigate the molecular signaling mechanism and in vivo tumorigenic ability of Mfn2 regulating the biological behavior of cervical cancer cellsObjective: To investigate the molecular signaling mechanism and in vivo tumorigenesis of Mfn2-regulated cervical cancer cell biological behavior.Methods: The morphology and quantity of cytoskeleton after silencing Mfn2 were detected by phalloidin staining.Western blotting was used to detect the expression of Ras family protein Rho A and downstream ROCK kinase.Western blotting was used to detect the changes of key proteins in IL-6 / STAT3 signaling pathway after silencing Mfn2.Results: After silencing Mfn2,the number of cytoskeleton decreased,the number of pseudopodia decreased and the athletic ability decreased.The expression of Rho A and ROCK kinase in Ras family decreased(P <0.05)after silencing Mfn2.After silencing Mfn2,the key protein expression levels of the IL-6 / STAT3 signaling pathway were correspondingly reduced.(P <0.05).The expression of Mfn2,p-STAT3 and Rho family protein was decreased in the tumor tissue of mice treated with Mfn2.Conclusion: Mfn2 can regulate the cytoskeleton of cervical cancer cells,and through IL-6 / STAT3 signaling pathway and affect the ability of cervical cancer cells.Silencing of Mfn2 expression on cervical cancer cells also have a certain degree of inhibition of tumorigenesis.