shRNA-induced knockdown of the SPERT gene inhibits proliferation and promotes apoptosis of human colorectal cancer RKO cells

Abstract

Colorectal cancer is the third most common type of cancer and the fourth leading cause of cancer-related deaths worldwide. Although several genes have been identified to contribute to the pathogenesis of colorectal cancer, there are still many genes with unidentified functions in colorectal cancer. This study aimed to investigate the effect of shRNA-induced knockdown of the SPERT gene on the proliferation and apoptosis of human colorectal cancer RKO cells. SPERT was screened based on the TCGA dataset, and SPERT expression, cell growth, proliferation and apoptosis were detected in shSPERT- and shCtrl-transfected RKO cells. In addition, the SPERT-related biological pathways were detected using a PathScan® Signaling Antibody Array Kit. We detected lower SPERT expression in shSPERT-transfected RKO cells than in shCtrl-transfected cells at both the translational and transcriptional levels (P<0.05), and an MTT assay revealed a clear-cut decrease in the proliferation of shSPERT-transfected RKO cells relative to shCtrl-transfected RKO cells (P<0.01). A Caspase-Glo® 3/7 assay detected an increase in the caspase-3/7 activity and the number of apoptotic cells in the shSPERT-transfected RKO cells than in the shCtrl-transfected cells (P<0.01), and flow cytometry detected a higher apoptotic rate in the shSPERT-transfected RKO cells than in the shCtrl-transfected cells (20.65±0.26 vs. 5.93±0.06%, respectively, P<0.01). Elevated levels of phosphorylated p44/42 MAPK (ERK1/2), Akt, Bad, HSP27, p38 MARK and Chk2, and elevated PARP and caspase-3 expression levels were detected in shSPERT-transfected RKO cells compared with the shCtrl-transfected cells (P<0.05). The results of the current study demonstrated that knockdown of SPERT suppresses colorectal cancer cell growth and promotes apoptosis. SPERT may serve as an oncogene and may be a potential target for the treatment of colorectal cancer.