Expression And Significance Of PKHD1 Gene In Colorectal

Objective:To study the relationship between PKHD1 gene expression and colorectal cancer.To provide a new experimental basis for the molecular mechanism of colorectal cancer.Methods:The expression of PKHD1 gene in colorectal tissues and cell lines was detected by immunoblotting.sh RNA targeting PKHD1 was built in p LKO.1-sh RNA-PKHD1 l lentiviral vector and transfected into normal colonic mucosa epithelial cell line(NCM460).The cells were selected using G418 and the p LKO.1-sh RNA-PKHD1 successful transfeced in NCM460 cells were observed by fluorescence microscopic,double digestion and DNA sequencing.In order to make sure that the p LKO.1-sh3RNA-PKHD1-NCM460 was constructed successfully.We detected the expression level of PKHD1 by Quantitative Real-time RT-PCR(q RT-PCR)and Western Blotting.The effects of PKHD1 knock-down on cells cell invasion and metastasis of NCM460 cells were evaluated by Transwell,.Results: The results of Western Blotting revealed that the expression of PKHD1 gene was low in colorectal cancer tissue and cell lines.The lentiviral vector targeting PKHD1 gene was successfully constructed and efficiently infected into NCM460 ce1 ls.The results of q RT-PCR and Western Blotting revealed that sh RNA against PKHD1 significantly decreased the m RNA and protein expressions of PKHD1.The NCM460 cell line of knock-down PKHD1 was successfully established.Transwell experiment showed that cell migration were(232.67 ± 12.36)in experimental group,which was much higher than that in the bland load transfection group and control group(58.22 ± 9.79 and 66.14 ±9.28).Transwell experiment showed that cell invasion were(182.53±11.36)in experimental group,which was much higher than that in the bland load transfection group and control group(46.73±7.93 and 47.23±7.36).Transwell results showed that the migration and invasion of NCM460 cells were significantly increased after low expression of PKHD1 gene.Conclusions: 1.The expression of PKHD1 gene was low in colorectal cancer tissue and cell lines;2.Knockdown of PKHD1 can enhance the abilities of NCM460 cells in migration and invasion.