Retrospective Study
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World J Gastroenterol. Jun 14, 2014; 20(22): 6974-6980
Published online Jun 14, 2014. doi: 10.3748/wjg.v20.i22.6974
Influence of adriamycin on changes in Nanog, Oct-4, Sox2, ARID1 and Wnt5b expression in liver cancer stem cells
Ding Sun, Lei Qin, Yang Xu, Jian-Xia Liu, Li-Ping Tian, Hai-Xin Qian
Ding Sun, Lei Qin, Yang Xu, Jian-Xia Liu, Li-Ping Tian, Hai-Xin Qian, Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China
Author contributions: Sun D and Qin L performed the majority of the experiments; Xu Y, Liu JX and Tian LP provided vital reagents and analytical tools, and were involved in editing the manuscript; Qian HX collected all materials and provided financial support for this work; Sun D designed the study and wrote the manuscript.
Supported by National Natural Science Foundation, No. 81372317
Correspondence to: Hai-Xin Qian, Chief Physician, Department of General Surgery, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou 215006, Jiangsu Province, China. sunding0126@gmail.com
Telephone: +86-512-65223637 Fax: +86-512-65223637
Received: March 8, 2014
Revised: April 28, 2014
Accepted: May 19, 2014
Published online: June 14, 2014
Processing time: 100 Days and 9.1 Hours
Abstract

AIM: To determine the influence of Adriamycin (ADM) on the changes in Nanog, Oct4, Sox2, as well as, in ARID1 and Wnt5b expression in liver cancer stem cells.

METHODS: The MHCC97-L and HCCLM3 liver cancer cell lines were selected as the cell models in this study, and were routinely cultured. The 50% lethal dose (LD50) in the cell lines was detected by the MTT assay. Expression changes in liver cancer stem cell related genes (Nanog, Oct-4, Sox2, ARID1, and Wnt5b) were detected by western blot following treatment with ADM (LD50).

RESULTS: The LD50 of ADM in MHCC97-L cells was lower than that in HCCLM3 cells (0.4123 ± 0.0236 μmol/L vs 0.5259 ± 0.0125 μmol/L, P < 0.05). Wnt5b and Nanog were expressed in both MHCC97-L and HCCLM3 cells, while only Sox2 was expressed in HCCLM3 cells. However, neither ARID1A nor Oct4 was detected in these two cell lines. Genes, related to the stem cells, showed different expression in liver cancer cells with different metastatic potential following treatment with ADM (LD50). Wnt5b protein increased gradually within 4 h of ADM (LD50) treatment, while Nanog decreased (P < 0.05). After 12 h, Wnt5b decreased gradually, while Nanog increased steadily (P < 0.05). In addition, only Sox2 was expressed in HCCLM3 cells with high metastatic potential following ADM (LD50) treatment. The expression of Sox2 increased gradually with ADM (LD50) in HCCLM3 cells (P < 0.05).

CONCLUSION: ADM increased the death rate of MHCC97-L and HCCLM3 cells, while the growth suppressive effect of ADM was higher in MHCC97-L cells than in HCCLM3 cells.

Keywords: Liver cancer cell; Adriamycin; Stem cell related gene; Western blot; Liver cancer stem cell

Core tip: This study, comprehensively and systematically reports the gene expression of Nanog, Wnt5b, Oct4, and Sox2 and ARID1A, in two hepatoma cell lines. This was performed to explore the molecular mechanism of the recurrence and metastasis of hepatocellular carcinoma, and to detect accurate markers and targets for stem cell-targeted therapeutic interventions. The results showed that Adriamycin (ADM) increased the death rate of MHCC97-L and HCCLM3 cells, while the growth suppressive effect of ADM was higher in MHCC97-L cells than in HCCLM3 cells.