全反式維甲酸對HepG2肝癌干細胞增殖能力的影響

， ， ， ，

1.西安市第四醫院普外科，陜西 西安 710004；2.西安交通大學第一附屬醫院血液內科

【Abstract】ObjectiveTo investigate the effect of all-transretinoic acid (ATRA) on the multiplication capacity of HepG2 hepatocarcinoma stem cells.MethodsHepG2 liver cancer cells were inoculated on 96-well plates and hepatocarcinoma stem cells were cultured and identificated, which based on the principles randomly assigned into the ATRA group and control group, each group of 48 hole, ATRA group was given 5 μmol/L ATRA and the control group did not make any processing.ResultsExpressions of Cyclin D and STAT3 by Western blotting method in the control group showed a trend of rising, those in ATRA group reduced after rising first, expressions of Cyclin D and STAT3 in the 5, 7, 14 days in the ATRA group were significantly lower than those in the control group, the difference was statistically significant (P<0.05). G0/G1cell proportion in the control group showed no significant change, that in the ATRA group showed a gradual upward trend, G0/G1cell proportion in the 5, 7, 14 days in the ATRA group were significantly higher than those in the control group, the difference was statistically significant (P<0.05). The stem cell A570 value in the control group showed a gradual upward trend, that in the ATRA group showed a gradual decline, the stem cell A570 value in the 5, 7, 14 days in the ATRA group were significantly lower than those in the control group, the difference was statistically significant (P<0.05).ConclusionATRA can effectively inhibit the proliferation of HepG2 hepatocarinoma stem cells, and its mechanism may be related to the reduction of gene protein expressions of Cyclin D and STAT3 in stem cells and blocking stem cells in G0/G1cell cycle.

【Keywords】 All-transretinoic acid; HepG2 hepatocarcinoma stem cells; Multiplication capacity; Cell cycle

肝癌是臨床上常見的消化系統惡性腫瘤之一，具有發病率高、死亡率高等特點，其常用的治療方法為外科手術治療，通過切除癌灶組織可有效控制患者的病情，但多數患者被發現時已處于中晚期階段，手術治療療效欠佳，化療等其他治療成為極為重要的治療方法。而有研究[2-3]認為，腫瘤組織中存在一類具有干細胞性狀的腫瘤細胞即腫瘤干細胞，其可導致腫瘤細胞出現化療耐藥，而致使療效欠佳，故如何提高患者的治療療效具有重要的臨床意義。有研究顯示，全反式維甲酸(all-transretinoic acid，ATRA)是臨床上常用的一種分化劑，對胚胎發育過程中細胞分化，機體生長、發育均具有重要調節作用，已逐漸被應用于腫瘤治療中，且具有良好的臨床療效，但其對肝癌干細胞增殖能力的抑制作用的報道還不多見。對此，本研究通過在肝癌細胞系HepG2中應用ATRA進行實驗，探討其對腫瘤干細胞增殖能力的影響，為臨床合理應用抗腫瘤藥物提供參考，現報道如下。

1 材料與方法

1.1主要材料和儀器肝癌細胞系HepG2細胞株(中國科學院昆明動物研究所)，DMEM培養基(中國上海GIBICO公司)，PBS磷酸鹽緩沖液(北京Solarbio公司)，雙抗(青霉素/鏈霉素)(中國上海Hyclone公司)，ATRA、二甲基亞砜(DMSO)和四甲基偶氮唑藍(MTT)(美國Sigma公司)，質量濃度為2.5 g/L的胰蛋白酶(中國上海Hyclone公司)，細胞因子EGF、FGF2、B27(德國Miltenyi公司)，新生胎牛血清(杭州四季青生物工程材料有限公司)，鼠抗人Cyclin D和STAT3抗體(上……