Tianjin Med J ›› 2015, Vol. 43 ›› Issue (12): 1368-1372.doi: 10.11958/j.issn.0253-9896.2015.12.007

• Cell and Molecular Biology • Previous Articles     Next Articles

Role of TNF-α in promoting migration and invasion of colon cancer cells

ZHAO Peng, CHEN Junmao, CAO Wenbin, YANG Guanghua, YU Xiangyang, LIU Chunhui, ZHENG Yang   

  1. Department of General Surgery, North China University of Science and Technology Affiliated Hospital, Tangshan 063000, China
  • Received:2015-05-13 Revised:2015-09-15 Published:2015-12-15 Online:2015-12-11

Abstract: Objective To explore the effect of TNF-α on expression of TROP-2 and to explore the role of TROP-2 in the metastasis and invasion of colon cancer HCT-116 cells. Methods HCT-116 cells were cultured and treated with 0, 10, 20, 30, 50, 100 and 200 μg/L TNF-α. Cell viability was assessed by MTT. The expression of TROP-2 was determined by western blot. The effects of 20 μg/L TNF-α on cell migration and invasion were investigated by wound healing assay and Transwell method. Small interfering RNA (siRNA) was used to knock down endogenous TROP-2 expression. The transcrip⁃ tion and translation levels of TROP-2 were detected by qPCR and Western blot respectively. The migratory and invasive ca⁃ pability of HCT-116 cells transfected with TROP-2 siRNA were checked by wound healing assay and Transwell method re⁃ spectively. Results There is no significant change of cell viability between HCT-116 cells treated with 0,10, 20, 30 and 50 μg/L TNF-α, but cell viability of HCT-116 decreased significantly with treatment of 100 μg/L and 200 μg/L TNF-α. Low concentration of TNF-α (≤50 μg/L) led to increase of TROP-2 protein expression that peaks when 20 μg/L TNF-α was add⁃ ed. High concentration of TNF-α (100, 200 μg/L) result in decrease of TROP-2 protein. TROP-2 siRNA significantly downregulated the expression of TROP-2 at both mRNA and protein levels in colon cancer HCT-116 cells. Compared with con⁃ trol group, silencing TROP-2 by TROP-2 siRNA inhibited the migratory and invasive capability of HCT-116 cells. Wound healing rate and the number of transwell cell both decreased in siRNA group compared with those of control group (P < 0.05). Conclusion The mechanism that low concentration of TNF-α promoted HCT-116 cells migration and invasion might be through up-regulating the expression of TROP-2.

Key words: colonic neoplasms, tumor necrosis factor-alpha, neoplasm invasiveness, cell movement, TROP-2