Effect of small interfering RNA silencing vivo hydrocarbon receptor nuclear transporter gene on the colon cancer HCT8 cell growth

Abstract

Objective To observe the influence of small interfering RNA (siRNA) silencing Aryl hydrocarbon receptor nuclear transporter (ARNT) gene on colon cancer HCT8cell growth and vivo tumorigenicity. Methods Colon carcinoma cell HCT8 was transfected with ARNT-short hairpin RNA (shRNA) lentivirus. Rats subcutaneous tumor model were established. We used polymerase chain reaction (PCR) to assay the ARNT mRNA expression of HCT8 cell, Western blotting to assay ARNT protein level of HCT8 cells, cell counting kit-8 (CCK-8) to assay the proliferation of HCT8 cell, flow cytometry to assay the apoptosis of HCT8 cell. We weighed the tumor weight and measured tumor volume at one month after modeled. Results The ARNT mRNA relative expression of siRNA-ARNT group (0.489±0.057) was significantly lower than that of siRNA-NC group (0.921±0.102) and control group (1.000±0.000) (P=0.000). The ARNT protein expression level of siRNA-ARNT group (0.478±0.025) was significantly lower than that of siRNA-NC group (0.818±0.017) and control group (0.827±0.013) (P=0.000). The 24, 48, 72 h Cells OD value of siRNA-ARNT group (0.824±0.121, 1.511±0.142, 1.602±0.154) were higher than that of siRNA-NC group (0.512±0.107, 1.012±0.123, 1.121±0.128) and control group (0.503±0.956, 0.989±0.121, 1.116±0.131) (P=0.000、0.000、0.000). The cell apoptosis rate of siRNA-ARNT group [(55.13±2.78)%] was higher than that of control group [(4.75±0.68)%, P=0.000], lower than that of siRNA-NC group [(68.32±3.12)%, P=0.000], the cell apoptosis rate of siRNA-NC group was higher than that of control group (P=0.000). Rat tumor weight [(0.551±0.171) g] and volume (0.557±0.153) cm3 of siRNA-ARNT group were higher than that of siRNA-NC group [(0.351±0.082) g, (0.414±0.071) cm3] and control group [(0.325±0.111) g, (0.396±0.063) cm3,P=0.000, 0.000]. Conclusion Silencing ARNT gene can promote the proliferation of colon cancer HCT 8 cell, inhibite the apoptosis of HCT8 and promote vivo tumor growth. ARNT gene may be colon cancer suppressor gene. Key words: Aryl hydrocarbon receptor nuclear transporter; HCT8 cell; RNA interference; Proliferation; Apoptosis