Effect Of RACK1 On EMT-related Factor Expression In SiHa Cell Of Cervical Cancer

The incidence of cervical cancer ranks fourth among female malignant tumors in the world.The health of women of different ages suffers its harm,and the age of onset is getting smaller,which seriously affects the quality of life of young women.Further progression,metastasis,and malignant transformation of cervical cancer are important factors that cause death.It is particularly important to study the mechanism of occurrence and development of cervical cancer and block its further invasion and metastasis.Receptor for activated C kinase 1(RACK1)is a structure built many intracellular signaling pathways involved in protein,more RACK1 proteins in combination with surface can be combined with a variety of factors,regulating tumor cell migration,invasion,raised colonization and apoptosis resistance,etc,the function of regulating tumor progression.There have been reports of expression and significance in a variety of tumors.Epithelial-mesenchymal transition(EMT)in a variety of epithelial sex play an important role in development and incidence of malignant tumor,EMT refers to the physiological and pathological epithelial cells in specific cases to mesenchymal cells,cancer cells after EMT adhesive decline or disappear,cell movement ability strong rights,thereby gaining breakthrough basement membrane,the ability to invade surrounding tissues,enter the lymphatic or blood vessel to distant organs,studies confirm that EMT can promote the invasion and metastasis of cervical cancer cells,strengthen the malignant degree of cervical cancer.The expression of RACK1 can reduce the expression of e-cadherin and increase the expression of waveform protein,which is consistent with the occurrence of EMT.In this experiment,SiHa cell of cervical cancer were studied and the recombinant plasmid RACK1 was transfected into SiHa cell by liposome method to obtain SiHa cell over-expressing RACK1.The EMT-related protein E-cadherin in SiHa cell overexpressing RACK1 was analyzed.The expression of vimentin was analyzed,and the relationship between overexpressed RACK1 and cervical cancer EMT was analyzed.Themolecular mechanism of invasion,metastasis and malignant transformation of cervical cancer was studied to provide basis for the diagnosis and treatment of cervical cancer.Objective: The expression of e-cadherin and vimentin in SiHa cell of cervical cancer can be affected by the expression of RACK1,so as to induce the transformation of epithelium and promote its proliferation and migration.Methods: 1.Cervical cancer SiHa cell culture and RACK1 plasmid transfection;2.The expression level of RACK1,e-cadherin,and vimentin m RNA of the cells(recombinant plasmid RACK1 experimental group;The negative control group of transfected empty plasmid;the blank control group without transfection)were detected by RT-qPCR.3.Western blot method was used to detect the protein expression level of e-cadherin and vimentin in each group of cells.4.All data use SPSS16.0 statistical analysis software,set the differences between the single factor analysis of variance(SNK method)is adopted to improve the statistical analysis,the differences in the group using single factor analysis of variance method(LSD)statistical analysis;All data were expressed in the form of mean plus or minus standard deviation(plus or minus S),which was considered statistically significant by P < 0.05.Results: 1.The result of RT-qPCR test showed that the expression level of RACK1 m RNA in the experimental group cells(1.25±0.59)was higher than that in the negative control group(0.71±0.12)and the blank control group(0.66±0.07),the difference was statistically significant(p<0.01).2.RT-qPCR method measured the expression level of vimentin m RNA in the experimental group cells(3.42±1.04)higher than that in the negative control group(1.13±0.19)and the blank control group(1.06±0.13),and the m RNA expression level of e-cadherin(0.38±0.09)was lower than that in the negative control group(1.08±0.20)and the blank control group(1.13±0.19),the difference was both statistically significant(p<0.01).3.Western blot method measured the protein expression level of RACK1 in the experimental group cells(0.94±0.08)above the negative control group(0.74±0.07)and the blank control group(0.72±0.09);the protein expression level of vimentin in the experimental group cells(1.13±0.12)above the negative control group(0.65±0.10)and the blank control group(0.58±0.07),while the expression level of e-cadherin protein(0.53±0.11)was lower than that in the negative control group(1.06±0.10)and the blank control group(1.13±0.11),the difference was both statistically significant(p<0.01).Conclusion: 1.Recombinant plasmid RACK1 successfully transfects the cervical cancer SiHa cell.2.Overexpression of RACK1 in promoting the expression of vimentin in SiHa cell of cervical cancer,inhibiting the expression of e-cadherin,in order to promote EMT of cervical cancer SiHa cell.