Effects of endoplasmic reticulum oxidoreduclin 1α knockdown on proliferation, apoptosis, migration and autophagy of colon cancer cells / 肿瘤

ABSTRACT

Objective: To investigate the effects of knocking down endoplasmic reticulum oxidoreduclin 1α (ERO 1α) gene expression on the proliferation, apoptosis, migration and autophagy of colon cancer cells, as wells as the possible mechanism. Methods: The Cancer Genome Atlas (TCGA) database was used to verify the expression of ERO1α mRNA in colon cancer tissues. The expression of ERO1α protein in colon cancer cell lines SW480, RKO, SW620, HCT116 and HT29 was detected by Western blotting. The recombinant lentiviruses carrying ERO1α-shRNA (shERO1α) and the control (shCtrl) were constructed and infected into colon cancer RKO cells, respectively. The fluorescence microscopy was used to evaluate the infection efficiency of lentiviruses in the two groups after 72 h. The expression level of ERO1α protein in RKO cells after infection was detected by Western blotting. The apoptosis of RKO cells was assessed by flow cytometry, and the expression levels of apoptosis-related molecules were detected by Western blotting. The proliferation of RKO cells was detected by Cellomics cell counter, and the cell cloning formation ability was detected by plate cloning formation assay. The migration ability of RKO cells was detected by Transwell chamber assay. Finally, the expression of key molecules in autophagy was detected by Western blotting. Results: The expression level of ERO1α mRNA in colon cancer tissues was significantly higher than that in the adjacent tissues (P < 0.001). The expression level of ERO1α protein in RKO cells was highest among the detected colon cancer cells. The recombinant lentivirus shERO1α was constructed successfully, and its infection efficiency into RKO cells was more than 80%. Compared with the control group, shERO1α could effectively knock down the expression level of ERO 1α gene (P < 0.01). ERO1α knockdown promoted cell apoptosis (P < 0.05), increased Caspase-3 expression (P < 0.01), and decreased Bcl-2 expression (P < 0.01). The proliferation, cloning formation and migration abilities of RKO cells were decreased after ERO1α knockdown (all P < 0.01). The expressions of autophogy-related molecules LC3 and Beclin 1 were increased (P < 0.05), and the expression of P62 was decreased (P < 0.01) in ERO1α knockdown cells. Conclusion: The ERO1α is highly expressed in colon cancer tissues and cell lines. Knockdown of ERO 1α gene can promote apoptosis, inhibit proliferation and migration, promote autophagy of colon cancer cells, suggesting that ERO1α plays an important role in the malignant process of colon cancer.