A Study On The Promotion Of Proliferation,Invasion And Migration Of Cervical Cancer Cells By HOXD3

Objective:The study explores the role of HOXD3 in promoting proliferation,invasion and migration of cervical cancer cells,and provides a new target for the treatment of cervical cancer.Methods:1.The relationship between HOXD3 expression and clinical prognosis of cervical cancer patients was searched through TCGA database;2.Immunohistochemical method was used to detect the expression level of HOXD3 in 77 cases of cervical squamous cell carcinoma,CIN,adjacent normal squamous cell cervical tissue and normal cervical tissue(normal squamous cell cervical tissue specimens were collected from patients with benign gynecological diseases and negative cervical cancer screening in the same period);3.The correlation between abnormal expression of HOXD3 and clinicopath-ological parameters of cervical cancer patients was analyzed;4.The expression levels of HOXD3 in He La and Si Ha cervical cancer cell lines were detected by Western Blot.Si Ha cells with low expression of HOXD3(si RNA-HOXD3)were obtained by si RNA knockdown transfection.The efficiency of HOXD3 transfection was detected by Western Blot and subsequent cell function experiments were performed;5.The role of HOXD3 in the proliferation,invasion and migration of cervical cancer cells was verified by cell malignant biological experiments.Results:1.The results in the TCGA database showed that the recurrence free survival(RFS)of patients with high HOXD3 expression was significantly shorter than that of patients with low HOXD3 expression,and the Logrank test was statistically significant(P= 0.031,HR = 2.33),The results suggest that HOXD3 may be a poor prognostic factor in patients with cervical cancer;2.The results of immunohistochemistry showed that most of the staining of HOXD3 protein was localized in the cytoplasm.The high expression of HOXD3 gradually deepened and the expression gradually strengthened with the deterioration of CIN,especially in the middle and late stage(II-III).The positive expression rate of HOXD3 in cervical squamous cell carcinoma was significantly higher than that in adjacent normal squamous cell cervical tissue,normal cervical tissue and CIN,with statistical significance(Z = 36.631,P < 0.001).The difference between normal cervical tissue,CIN and adjacent normal squamous cell cervical tissue and advanced stage was the most significant,with statistical significance(P = 0.031,P = 0.01,P <0.0001);3.In cervical squamous cell carcinoma tissues,using non-parametric test-k independent samples(Kruskal-Wallis)test,it was found that the high expression of HOXD3 was not statistically significant with the patient ’s age(P= 0.093),interstitial infiltration depth(P= 0.814),vascular tumor thrombus(P= 0.188),Ki-67 percentage(P= 0.406),CK high and low molecular detection(P= 0.9),but was statistically significant with 2018 FIGO clinical stage and lymph node metastasis(P= 0.043).The results of SPearman correlation analysis showed that the high expression of HOXD3 was positively correlated with lymph node metastasis and2018 FIGO clinical stage(rs = 0.314,P= 0.012).There was statistical significance(rs = 0.257,P= 0.042);4.The results of Western-blot showed that HOXD3 was significantly higher in cervical squamous cell carcinoma Si Ha cell line than in cervical adenocarcinoma He La cell line(P = 0.0445).After transfection of si RNA-HOXD3,the expression of HOXD3 protein in Si Ha cell line was significantly knocked down(P = 0.0358),which was used for subsequent cell function experiments;5.The results of cell function experiments showed that knockdown of HOXD3 could significantly inhibit the proliferation,invasion and migration of cervical cancer Si Ha cell line(scratch test P= 0.0218;invasion experiment P= 0.0411;migration experiment P= 0.003;CCK8 experiment 96 h P= 0.0011).Conclusion:1.HOXD3 is highly expressed in cervical cancer tissues and is positively2.correlated with 2018 FIGO clinical stage and lymph node metastasis of cervical cancer.3.HOXD3 is highly expressed in cervical cancer Si Ha cell line and can participate in the proliferation,invasion and migration of cervical cancer cells.Knockdown of HOXD3 expression can significantly inhibit the proliferation,invasion and migration of cervical cancer cells.