Mechanism research on apoptosis of hepatocellular carcinoma induced by sodium bicarbonate
ZHANG Yan1 QIAO Luxin2 ZENG Jing2 XU Jun1 SHI Ying2▲
1.Beijing Zhongguancun Middle School, Beijing 100086, China;
2.Beijing You′an Hospital, Capital Medical University Beijing Institute of Hepatology, Beijing 100069, China
Abstract：Objective To explore the apoptotic mechanism of hepatocellular carcinoma (HCC) induced by sodium bicarbonate (SB). Methods HepG2 cells were implanted the subcutaneous of 5 athymic mice (7-week age) to form hepatocarcinoma mouse model and the growth and necrosis of hepatocellular carcinoma was observed under the condition of local mass injection of 5% SB. The apoptosis of hepatocellular carcinoma cell line HepG2 was observed by Calcein/PI staining and flow cytometry at SB exposure and different pH gradients. The expression of Bax/bcl2 were also detected in HepG2 cells under SB exposure by real-time PCR and western blotting. Results Animal in vivo studies revealed that 5% SB local injection for 4 weeks could inhibit tumor growth and cause tumor necrosis in 3 of 5 hepatocarcinoma mice. One hepatocarcinoma mice died and another mice had an enlarged tumor. 5% SB was diluted to the volume ratio 3/4, 1/2, 1/4, 1/8, 1/16 and negative control by DMEM and Calcein/PI staining in vitro showed that HepG2 apoptosis mice decreased in turn. Flow cytometry showed the similar results in 5% SB volume ratio: 1/2, 1/4, 1/8 and 1/16. The medium DMEM was titrated to pH6.0, pH6.25, pH6.5, pH6.75, pH7.0, pH7.25, pH7.5, pH7.75 and pH8.0. The lowest rate of HepG2 apoptosis was at pH7.0 and pH7.25. As the increasing of acidity or alkalinity, the apoptosis rate of HepG2 increased gradually. Flow cytometry also showed the similar results. Compared with control, Bax mRNA and protein expression in SB exposed HepG2 cell were not changed, but bcl2 mRNA and protein expression were decreased. Conclusion SB can induce HCC apoptosis by changing the internal environmental pH and inhibiting the expression of apoptotic protein blc2.
[1] 陈万青，张思维，曾红艳，等.中国2010年恶性肿瘤发病与死亡[J].中国肿瘤，2014，23（1）：1-10.
[2] 王安娜，闫晓东，封晓昆，等.乙肝病毒核心启动子区和前C区突变与肝细胞自噬的关系[J].北京医学，2014，36（9）：756-759，799.
[3] 贾士杰，范慧敏，刘伟，等.2002～2011年中国恶性肿瘤死亡率水平及变化趋势[J].中国肿瘤，2014，23（12）：999-1004.
[4] 樊嘉，王征.肝癌外科治疗的进展[J].实用医院临床杂志，2011，8（1）：16-19.
[5] 刘秀红，赵一鸣，赵晓飞，等.肝细胞癌诊断与治疗研究进展[J].中国肝脏病杂志：电子版，2017，9（2）：20-25.
[6] 孙惠川，王征，周俭，等.2017年版中国肝癌诊断和治疗规范解读[J].实用肿瘤杂志，2018，33（1）：1-3.
[7] Chao M，Wu H，Jin K，et al. A nonrandomized cohort and a randomized study of local control of large hepatocarcinoma by targeting intratumoral lactic acidosis [J]. Elife，2016，5. doi:10.7554/eLife.15691
[8] Robey IF，Baggett BK，Kirkpatrick ND，et al. Bicarbonate increases tumor pH and inhibits spontaneous metastases [J]. Cancer Res，2009，69（6）：2260-2268.
[9] Zhang Y，Shi Y，Qiao L，et al. Sigma-1 receptor agonists provide neuroprotection against gp120 via a change in bcl-2 expression in mouse neuronal cultures [J]. Brain Res，2012，1431：13-22.
[10] 左婷婷，郑荣寿，曾红梅，等.中国肝癌发病状况与趋势分析[J].中华肿瘤杂志，2015（9）：691-696.
[11] 吕桂帅，陈磊，王红阳.我国肝癌研究的现状与前景[J].生命科学，2015，27（3）：237-248.
[12] 黄洁夫，李绍强，梁力建.肝动脉化疗栓塞在原发性肝癌治疗中的地位和作用[J].中华肝胆外科杂志，2000，6（1）：3-6.
[13] Al-Jaghbeer M，Kellum JA. Acid–base disturbances in intensive care patients：etiology，pathophysiology and treatment [J]. Nephrology Dialysis Transplantation，2014，30（7）：1104-1111.
[14] 王岚，尤琳浩，常彦忠.人体维持酸碱平衡的机制[J].生物学通报，2013，48（2）：1-2.
[15] Estrella V，Chen T，Lloyd M，et al. Acidity generated by the tumor microenvironment drives local invasion [J]. Cancer Res，2013，73（5）：1524-1535.
[16] Silva AS，Yunes JA，Gillies RJ，et al. The potential role of systemic buffers in reducing intratumoral extracellular pH and acid-mediated invasion [J]. Cancer res，2009，69（6）：2677-2684.
2018, Vol. 15 
京公网安备 11010502046598号