KLF12 Regulates CDH11 Expression To Promote Endometrial Cancer Cells Proliferation

Objective: Endometrial cancer(EC)has become one of the most common malignant tumors of the female reproductive system in developing countries,with its local recurrence and distant metastasis after operation becoming the main problems endangering the quality of life of patients.Therefore,it is urgent to study the occurrence and development of endometrial cancer.In recent years,the molecular mechanism research of the occurrence and development of endometrial cancer has been conducted in our laboratory,and it has been confirmed that KLF12 regulates some biological behavior of endometrial cancer cells.However,how KLF12 regulates the target genes involved in the growth regulation of endometrial cancer has yet to be investigated.In this study,we detect the downstream target gene of KLF12,and explore how KLF12 participates in the growth regulation mechanism of endometrial cancer,which is of great significance to prevent and treat the recurrence of endometrial cancer after surgery.Methods: 1.Western blotting and q-PCR are adopted to detect the white analysis of normal endometrial cells and endometrial cancer cells to understand the expression of KLF12 in endometrial cancer cells;2.changes of KLF12 over-expressing related genes are analyzed in accordance with gene chip data,thus selecting specific target genes from related target genes;3.The difference of related target genes is analyzed by RT-q PCR technology,thus obtaining the relevant KLF12 m RNA expression level of target gene;4.KLF12 expression level in endometrial cancer cells and human normal endometrial cells is detected with western blotting and q-PCR detection,and expression of KLF12 related target gene protein levels are also analyzed;5.Western blotting and q-PCR are used to detect the silencing and overexpression of KLF12 affecting the expression level of KLF12 and its target genes in endometrial cancer cells.Silencing CDH11 affect the expression level of KLF12 and its target genes in endometrial cancer cells.Silencing the overexpression of CDH11 gene in KLF12 endometrial cancer cells result in the expression level of KLF12 and its target genes.6.Specific biological role of KLF12 and specific target genes in endometrial cancer cells is analyzed by CCK-8 experiment.Results:(1)Overexpression of KLF12 is found in endometrial cancer cells and normal endometrial cells Significant difference * P< 0.05.(2)The results of microarray analysis showed that the overexpression of KLF 12 resulted in the target gene differences of promoters with specific sequence bases as follows: ARHGDIB,BLVRA,CDH11,CDK2AP2,CLEC2 D,CTNNB1,FABP6,GP2,LRAP,MCM7,MST1,MT1 H,NET1,PAGE1,PGCP,RPL37,SCAMP1,TFF1,TMEM195,ZNF581.The promoters of the above genes are subject to KLF 12.Regulation.(3)We can get the target gene expression level of KLF12 through analysis of q RT-PCR.Overexpression of KLF12 leads to overexpression of downstream target genes CDH11,CAEC2 D,MST1,and overexpression of KLF12 reduces downstream target genes ARHGDIB,BLVRA,CDK2AP2,FABP6,MCM7,MT1 H,NET1,PAGE1.Overexpression of PGCP,RPL37,SCAMP1,ZNF581,KLF12 and downstream target genes CTNNB1 TMEM195 are not statistically significant;Overexpression of KLF12 regulated downstream target genes and gene chip detection are consistent.Overexpression of ARHGDIB,BLVRA,CDK2AP2,CLEC2 D,FABP6,MCM7,MT1 H,CDH11,MST1,KLF12 regulates downstream target genes.The inconsistencies with gene chip detection are: NET1,PAGE1,PGCP,SCAMP1,ZNF581.(4)Western blotting and q-PCR are utilized to detect the relative expression of KLF12 protein and CDH11 after silencing IK cells.The results show that the statistical difference exists between before and after silencing IK cells * P< 0.01.The relative expression of KLF12 and CDH11 in IK cells after silencing CDH11 gene is statistically different from that of CDH11 cells,* *(P< 0.01);Results also show that there is no statistical difference between KLF1 and KLF12,(P> 0.05.The relative expression of KLF12 and CDH11 protein in overexpressed IK cells is significantly different before and after treatment,* *(P< 0.01).Overexpressed IK cells silence CDH11 gene,and the relative expression of KLF12 and CDH11 protein before and after treatment compares with that of CDH11.No significant difference exists between KLF12 and KLF12 before and after treatment(P> 0.05).(5)There is a significant difference in the proliferation of CCK-8 cells between IK cells overexpressing KLF12 and IK cells silencing CDH11 overexpressing KLF12 * *(P< 0.01).Conclusion:(1)KLF12 is highly expressed in endometrial cancer cells.(2)KLF12 regulates target genes with specific promoters downstream(ARHGDIB,BLVRA,CDH11,CDK2AP2,CLEC2 D,CTNNB1,FABP6,GP2,LRAP,MCM7,MST1,MT1 H,NET1,PAGE1,PGCP,RPL37,SCAMP1,TFF1,TMEM195,ZNF58).(3)KLF12 upregulates the expression of CDH11 protein in endometrial cancer cells,as well as the biological behavior of CDH11 proliferation in endometrial cancer cells.