Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Trichlorophenyl-benzoxime induces apoptosis in human colon carcinoma cells via activation of mitochondria-dependent pathway

Zhongfu Xiao¹, Ke Zhang², Yun Huang², Yufan Zhou², Kuan Hu²

¹Department of Anesthesiology; ²Department of General Surgery, Xiang Ya Hospital, Central South University, Changsha, Hunan 410008, China.

For correspondence:- Kuan Hu Email: KennithKeithjzt@yahoo.com Tel:+8673184327191

Accepted: 25 June 2019 Published: 28 July 2019

Citation: Xiao Z, Zhang K, Huang Y, Zhou Y, Hu K. Trichlorophenyl-benzoxime induces apoptosis in human colon carcinoma cells via activation of mitochondria-dependent pathway. Trop J Pharm Res 2019; 18(7):1487-1492 doi: 10.4314/tjpr.v18i7.18

© 2019 The authors.
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Abstract

Purpose: To determine the apoptotic effect of trichlorophenyl-benzoxime (TCPB) on colorectal cancer (CRC) cells, and to elucidate the mechanism of action.

Methods: Colon carcinoma cell lines (DLD-1 and HT-29) were used in this study. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin at 37 I0;C in an atmosphere of 5 % CO₂ and 95 % air. When the cells attained 60 - 70 % confluency, they were treated with serum-free medium and graded concentrations of TCPB (1.0 – 6.0 μM) for 24 h. Cell viability and apoptosis were assessed using 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) and flow cytometric assays, respectively. Western blotting and 2', 7' dichlorofluorescein diacetate (DCFH DA) assays were used for the determination of expression levels of apoptotic proteins, and levels of reactive oxygen species (ROS), respectively.

Results: Treatment of DLD-1 and HT-29 cells with TCPB led to significant and dose-dependent reductions in their viability, as well as significant and dose-dependent increases in the number of apoptotic cells (p < 0.05). Treatment of HT-29 cells with TCPB led to significant increases in the population of cells in the G0/G1 phase, but significant reduction of cell proportion in S and G2/M phases (p < 0.05). It also significantly and dose-dependently upregulated the expressions of caspase-3 and bax, down-regulation of the expression of bcl-2 (p < 0.05). TCPB treatment upregulated the expressions of p53, cytochrome c (cyt c), procaspase-3, and procaspase-9, but down-regulated the expression of p-Akt dose-dependently (p < 0.05). The expression of Akt in HT-29 cells was not significantly affected by TCPB (p > 0.05). However, TCPB significantly enhanced the cleavage of PARP1, and significantly and dose-dependently increased the levels of ROS in HT-29 cells (p < 0.05).

Conclusion: These results suggest that TCPB exerts apoptotic effect on CRC cells via activation of mitochondria-dependent pathway, and thus can be suitably developed for the management of colon cancer.

Keywords: Colorectal cancer, Trichlorophenyl-benzoxime, Mitochondrial pathway, Apoptosis