CBX8 Activates JAK-STAT3 Pathway To Promote The Development Of Cervical Carcinoma

Objective:Cervical cancer is currently one of the most serious tumors that threaten women's health and has the highest degree of malignancy in the world.With the gradual development of medical technology in patients with cervical cancer in developed countries,the patient mortality rate has improved to a certain extent,but for the developing In countries or countries with poor economic or medical conditions,the incidence and mortality of cervical cancer remain high.Cervical cancer is a very serious malignant tumor in female tumors in our country,which seriously threatens its own health and the family's mental and economic situation.In China,the treatment of cervical cancer is mainly by surgical removal of tumor tissue and adjacent tissues,and combined with radiotherapy and chemotherapy before and after surgery.However,according to clinical data and postoperative follow-up of patients,most patients with recurrence of cervical cancer after surgery are facing resistance to chemotherapy drugs.Therefore,it is a huge difficulty for effective treatment of cervical cancer,so we can find new ones.Efficient treatment methods or new effective treatment drugs,for the adjuvant treatment of cervical cancer after surgery,delay or avoid the recurrence of cervical cancer and increase the prognosis of patients are currently urgently needed.This article aims to explore the impact of CBX8 in the Pc G protein family on the relevant biological processes of cervical cancer and its potential mechanism of action,to discover new molecularly targeted genes for the diagnosis and treatment of cervical cancer and to develop new anti-cervical cancer drugs.Clinically provide theoretical basis.Method:Part 1: First,by constructing CBX8 overexpression lentiviral vector and its empty vector,transfect human cervical cancer cell Si Ha cells,set the experimental group as the uninfected group(Control),infected with CBX8 overexpression empty vector lentiviral group(Empty vector)and infected CBX8 overexpression lentivirus group(OE-CBX8).CCK-8,scratch test and Transwell test were used to detect the effect of CBX8 on the cell viability,migration and invasion ability of each experimental group;secondly,we conducted a tumor formation experiment in nude mice.Finally,RT-PCR and Western blot methods were used to detect the expression of CBX8,STAT3,Bcl-2,p21 m RNA Part 2: First,by constructing CBX8 overexpression lentiviral vector and its empty vector,after transfecting human cervical cancer cell Si Ha cells,the experimental groups were set up to be infected with CBX8 overexpression empty vector lentivirus group(Empty vector)and infected CBX8 Overexpression of empty vector lentivirus + AG490 group(Empty vector + AG490),infection of CBX8 overexpression lentivirus group(OE-CBX8),infection of CBX8 overexpression lentivirus + AG490 group(OE-CBX8 + AG490).The effects of JAK / STAT3 pathway inhibitor AG490 on cell viability,migration and invasion ability of each experimental group were detected by CCK-8,scratch test and Transwell test.Then use RT-PCR and Western blot to detect the expression of STAT3,p-STAT3(ser727),Bcl-2,p21 m RNA and related proteins in each group of cells,to clarify the interaction between CBX8 and JAK / STAT3 pathway and the occurrence of cervical cancer The impact of development.Result:Part 1: The results showed that after overexpressing CBX8 transfected cervical cancer Si Ha cells,their cell proliferation and invasion and migration ability was significantly enhanced(P <0.01),the difference was statistically significant,the results proved that CBX8 overexpression can significantly enhance cervical cancer Cell viability,promote cell proliferation and metastasis.In order to prove the effect of CBX8 on cervical cancer in a complex environment in vivo,the results of nude mice tumorigenesis experiments proved that overexpressed CBX8 can promote the volume and weight of tumorigenesis in nude mice.To further study its role in the development of cervical cancer,RT-PCR and Western blot studies found that compared with the Control group,CBX8,STAT3,Bcl-2,p21 m RNA and CBX8,STAT3 in cervical cancer Si Ha cells of the Empty vector group,P-STAT3(ser727),Bcl-2,p21 protein expression levels have no significant changes,the difference is not statistically significant;compared with the Empty vector group,OE-CBX8 group cervical cancer Si Ha cells CBX8,STAT3,Bcl-2,p21 m RNA and CBX8,STAT3,p-STAT3(ser727),Bcl-2,p21 protein expression levels were significantly up-regulated(P <0.01),the difference was statistically significant.Part 2: The results show that compared with the Empty vector group,the cell viability and invasion and migration ability of the cervical cancer cell Si Ha in the Empty vector +AG490 group was significantly reduced(P <0.01),the difference was statistically significant;compared with the Empty vector group,The cell viability and invasion and migration ability of cervical cancer cell Si Ha in OE-CBX8 group were significantly enhanced(P <0.01),the difference was statistically significant.Compared with the OE-CBX8 group,the viability and invasion and migration ability of cervical cancer cells in the OE-CBX8 + AG490 group decreased significantly(P <0.01),and the difference was statistically significant.RT-PCR and Western blot were used to detect the expression of STAT3,Bcl-2,p21 m RNA and STAT3,p-STAT3(ser727),Bcl-2,p21 protein in the cells of each group.The results showed that compared with the Empty vector group,The expression levels of STAT3,Bcl-2,p21 m RNA and STAT3,p-STAT3(ser727),Bcl-2,p21 protein in cervical cancer Si Ha cells of Empty vector + AG490 group were significantly down-regulated(P <0.01),the difference was Statistical significance;compared with the Empty vector group,the expression levels of STAT3,Bcl-2,p21 m RNA and STAT3,p-STAT3(ser727),Bcl-2,p21 protein in cervical cancer Si Ha cells of the OE-CBX8 group were significantly Increased(P <0.01),the difference was statistically significant;compared with OE-CBX8 group,STAT3,Bcl-2,p21 m RNA and STAT3,p-STAT3(ser727)in cervical cancer cells of OE-CBX8 + AG490 group,The expression levels of Bcl-2 and p21 proteins were significantly down-regulated(P <0.01),and the difference was statistically significant.Conclusion:(1)CBX8 can promote the proliferation,migration and invasion of cervical cancer cells and play the role of oncogenes in the occurrence and development of cervical cancer.(2)CBX8 can up-regulate the expression levels of STAT3,Bcl-2,p21 m RNA and STAT3,p-STAT3(ser727)Bcl-2,p21 protein in cervical cancer cells related JAK /STAT3 signaling pathway(3)The JAK / STAT3 pathway inhibitor AG490 can significantly reverse the upregulation of cervical cancer cell proliferation,migration and invasion ability caused by CBX8 overexpression.(4)JAK / STAT3 pathway inhibitor AG490 can significantly reverse the expression of JAK / STAT3 pathway-related m RNA and protein in cervical cancer cells caused by CBX8 overexpression.(5)CBX8 promotes the occurrence and development of cervical cancer by activating the JAK / STAT3 signaling pathway.