Influence of long non-coding RNA SPRY4-IT1 on proliferation and apoptosis of colorectal cancer cells HT-29

ZHANG Debao; XING Chungen

doi:10.3969/j.issn.1002-266X.2017.13.001

Abstract：Objective To explore the influence and mechanism of long non-coding RNA (Lnc RNA) SPRY4-IT1 on proliferation and apoptosis of colorectal cancer cells HT-29.Methods Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of LncRNA SPRY4-IT1 in the colorectal cancer cell lines (HT-29, HCT-116, SW-480, SW-620, Caco-2) and the normal intestinal epithelial cell line FHC.We selected HT-29 cells with the relative highest expression of SPRY4-IT1 for this study.Lentivirus-mediated recombinant plasmid target SPRY4-IT1 was constructed and transfected into colorectal cancer HT-29 cells to establish the stably transfection cells of over-expression (up-regulation group) and interference (knock-down group).Meanwhile, the negative control group and blank control group were established.qRT-PCR was used to detect the expression of SPRY4-IT1 mRNA in HT-29 cells of each group.CCK-8 was applied to detect the proliferation of HT-29.cells Clone formation assay was used to test the colony forming efficincy of HT-29 cells.Flow cytometry was applied to detect the apoptosis and cell cycle of HT-29 cells.Western blotting was used to testify the expression of Bax and Bcl-2.Results The relative expression of LncRNA SPRY4-IT1 in colorectal cancer cell lines (HT-29, HCT-116, SW-480, SW-620, Caco-2) was significantly higher than that in the normal intestinal epithelial cell line FHC, and the expression of SPRY4-IT1 was the highest in HT-29 cells (all P<0.05).Compared with the negative control group and blank control group, the expression of SPRY4-IT1 mRNA, the proliferation and the colony forming efficincy of HT-29 cells in the up-regulation group were enhanced, and apoptosis rate was decreased (P<0.05 or P<0.01), and the opposite trend was observed in the knock-down group (P<0.05 or P<0.01).There was no significant difference in cell cycle of HT-29 cells among these groups (all P>0.05).Compared with the negative control group and blank control group, the expression of Bcl-2 was increased and Bax was decreased in HT-29 cells of the up-regulation group (P<0.05 or P<0.01), the expression of Bcl-2 was decreased and Bax was increased in HT-29 cells of the knock-down group (all P<0.01).Conclusion LncRNA SPRY4-IT1 may significantly accelerate proliferation and restrain apoptosis of colorectal cancer cell via promoting the expression of Bax and depressing the expression of Bcl-2.