Abstract:

Objective To investigate the effect of ITGB6 gene knockout on the biological behavior of human HT⁃29 colon cancer cells. Methods We constructed the ITGB6 knock⁃out HT⁃29 cells by CRISPR/Cas9 technolo⁃ gy（ITGB6⁃KO），which were defined as the knockout group. Meanwhile，the blank transfected cells were defined as the control group. Western blotting was used to detect the expression of ITGB6 in HT⁃29 cells. MTT assay，Plate colony formation assay and Transwell assay were used to detect the effects of ITGB6 knockout on the ability of proliferation，colony formation and invasion of HT⁃29 cells. Results Western blotting and DNA sequencing showed that CRISPR/Cas9 technology could effectively knock out ITGB6 gene. MTT assay，Plate colony formation assay and Transwell assay confirmed that knocking out ITGB6 gene could significantly inhibit the proliferative ability， colony formation ability and invasive ability of HT ⁃ 29 cells（P < 0.01）. Conclusion CRISPR/Cas9 technology can effectively establish a ITGB6 knock⁃out HT⁃29 cell line. After knocking out ITGB6 gene，the malignant biologi⁃ cal behavior of HT⁃29 cells is inhibited，therefore ITGB6 gene is expected to become a potential target for the treat⁃ ment of colon cancer.

Key words:

colon cancer, ITGB6, CRISPR/Cas9, colony formation, invasion