The Relationship Between The Expression Of COL3A1 In Cervical Cancer And HPV Infection And Its Effect On The Proliferation And Migration Of Siha Cells

Objective: 1)To analyze the correlation between collagen and HPV infection in cervical cancer;2)To explore the effect of COL3A1 on the proliferation and migration of cervical cancer cells.Methods: 1)Functional annotation,enrichment analysis and signal regulation network analysis of 9 kinds of collagen were performed by using DAVID,STRING and KOBAS databases to evaluate the relationship between collagen dysfunction and tumorigenesis.2)DNA was extracted from 38 cases of cervical cancer and 30 cases of non-cancerous cervical control tissues and q PCR was used to classify the HPV infection type of each sample.3)Total RNA was extracted from the above tissues and the transcription levels of 9 kinds of collagen in cervical cancer tissues were detected by q PCR method.4)30 cervical cancer tissue sections and 30 non-cancerous cervical tissue sections were selected to examine the expression level of COL3A1 protein by IHC.5)The expression of COL3A1 in three kinds of cervical cancer cells was detected by q PCR method.6)COL3A1 overexpression plasmid vector was constructed and transfected to Siha cells for establishing COL3A1 overexpression model;7)CCK8 assay was performed to detect the proliferation of Siha cells;8)The migration ability of Siha cells was detected using scratch test.Results: 1)The results of bioinformatics showed that among many biological processes and signal pathways involved in collagen,extracellular matrix organisation,cell adhesion,integrin binding,focal adhesion,human papillomavirus infection and PI3K-AKT signal pathway were closely related to the carcinogenisis of cervical cancer.2)HPV typing results: there were 41 HPV positive samples,including 36 cervical cancer tissues and 5 control tissues.HPV infection subtypes: 33 cases were positive for HPV16 alone,3 cases were positive for HPV16 with HPV18,and 5 cases were positive for HPV16 with HPV33/52/58/67.3)The results of q PCR analysis showed that the m RNA levels of COL1A2,COL3A1,COL6A1,COL6A3 and COL21A1 in cervical cancer tissues were significantly lower than those in the control group.4)Immunohistochemistry results showed that the expression of COL3A1 in cervical cancer was significantly down-regulated,which was consistent with the results of q PCR analysis.5)The expression level of COL3A1 in Siha and Hela cells was lower than that in C33 A cells,but there was no significant difference between Siha and Hela cells.6)After COL3A1 overexpression,the proliferation ability of Siha cells decreased significantly.7)The migration ability of Siha cells was significantly inhibited after overexpression of COL3A1.Conclusion: 1)Nine kinds of collagen down-regulated in cervical cancer are involved in a variety of tumor-related biological processes.2)The expression of COL1A2,COL3A1,COL6A1,COL6A3 and COL21A1 in HPV positive cervical cancer tissues is down-regulated.3)COL3A1 can inhibit the proliferation and migration of Siha cells and play an important role in the carcinogenisis of cervical cancer.