Original Article
Copyright ©2012 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jun 7, 2012; 18(21): 2640-2648
Published online Jun 7, 2012. doi: 10.3748/wjg.v18.i21.2640
Suppression of colorectal cancer metastasis by nigericin through inhibition of epithelial-mesenchymal transition
Hou-Min Zhou, Tao-Tao Dong, Lin-Lin Wang, Bo Feng, Hong-Chao Zhao, Xiu-Ke Fan, Min-Hua Zheng
Hou-Min Zhou, Bo Feng, Hong-Chao Zhao, Xiu-Ke Fan, Min-Hua Zheng, Department of General Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai Minimally Invasive Surgery Center, Shanghai 200025, China
Tao-Tao Dong, Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan 250012, Shandong Province, China
Lin-Lin Wang, Shanghai Institute of Medical Genetics, Shanghai Children’s Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China
Author contributions: Zhou HM and Dong TT contributed equally to this work; Zhou HM and Dong TT designed research; Feng B, Wang LL, Zhao HC and Fan XK provided analytical tools and were also involved in performed the research; Zhou HM wrote the manuscript; Zheng MH reviewed and modified the manuscript.
Supported by The National Natural Science Foundation, No. 30901424; and the Leading Medical Talent Foundation of Shanghai Municipality, No. 10XD1402700
Correspondence to: Dr. Min-Hua Zheng, Department of General Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai Minimally Invasive Surgery Center, No. 573 Xujiahui Road, Shanghai 200025, China. fengbo2022@163.com
Telephone: +86-21-64458887 Fax: +86-21-64458887
Received: December 6, 2011
Revised: February 1, 2012
Accepted: March 9, 2012
Published online: June 7, 2012
Abstract

AIM: To evaluate the effect of nigericin on colorectal cancer and to explore its possible mechanism.

METHODS: The human colorectal cancer (CRC) cell lines HT29 and SW480 were treated with nigericin or oxaliplatin under the conditions specified. Cell viability assay and invasion and metastasis assay were performed to evaluate the effect of nigericin on CRC cells. Sphere-forming assay and soft agar colony-forming assay were implemented to assess the action of nigericin on the cancer stem cell properties of CRC cells undergone epithelial-mesenchymal transition (EMT).

RESULTS: Compared with oxaliplatin, nigericin showed more toxicity for the HT29 cell line (IC50, 12.92 ± 0.25 μmol vs 37.68 ± 0.34 μmol). A similar result was also obtained with the SW116 cell line (IC50, 15.86 ± 0.18 μmol vs 41.02 ± 0.23 μmol). A Boyden chamber assay indicated that a significant decrease in the number of HT29 cells migrating through polyvinylidene fluoride membrane was observed in the nigericin-treated group, relative to the vehicle-treated group [11 ± 2 cells per high-power field (HPF) vs 19.33 ± 1.52 cells per HPF, P < 0.05]. Compared to the control group, the numbers of HT29 cells invading through the Matrigel-coated membrane also decreased in the nigericin-treated group (6.66 ± 1.52 cells per HPF vs 14.66 ± 1.52 cells per HPF, P < 0.05). Nigericin also reduced the proportion of CD133+ cells from 83.57% to 63.93%, relative to the control group (P < 0.05). Nigericin decreased the number of spheres relative to the control group (0.14 ± 0.01 vs 0.35 ± 0.01, P < 0.05), while oxaliplatin increased the number of spheres relative to the control group (0.75 ± 0.02 vs 0.35 ± 0.01; P < 0.05). Nigericin also showed a decreased ability to form colonies under anchorage-independent conditions in a standard soft agar assay after 14 d in culture, relative to the control group (1.66 ± 0.57 vs 7 ± 1.15, P < 0.05), whereas the colony numbers were higher in the oxaliplatin group relative to the vehicle-treated controls (14.33 ± 0.57 vs 7 ± 1.15, P < 0.05). We further detected the expression of E-cadherin and vimentin in cells treated with nigericin and oxaliplatin. The results showed that HT29 cells treated with nigericin induced an increase in E-cadherin expression and a decrease in the vimentin expression relative to vehicle controls. In contrast, oxaliplatin downregulated the expression of E-cadherin and upregulated the expression of vimentin in HT29 cells relative to vehicle controls.

CONCLUSION: This study demonstrated that nigericin could partly reverse the EMT process during cell invasion and metastasis.

Keywords: Colorectal cancer; Nigericin; Cancer invasion; Metastasis; Epithelial-mesenchymal transition; CD133; E-cadherin; Vimentin