Effect of Shenling Baizhu San Combined with Oxaliplatin on Proliferation and Apoptosis of Human Colon Cancer Cells

YU Wen-yan; WANG Hua-ying; WANG Guo-juan; LIU Jiang-yuan; LIU Zhi-ming; WEI Xiang-yan

doi:10.13422/j.cnki.syfjx.20181716

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Effect of Shenling Baizhu San Combined with Oxaliplatin on Proliferation and Apoptosis of Human Colon Cancer Cells

更新时间：2024-01-04

- Effect of Shenling Baizhu San Combined with Oxaliplatin on Proliferation and Apoptosis of Human Colon Cancer Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 18, Pages: 118-123(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20181716
  CLC： R285.5
- Published：2018
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YU Wen-yan, WANG Hua-ying, WANG Guo-juan, et al. Effect of Shenling Baizhu San Combined with Oxaliplatin on Proliferation and Apoptosis of Human Colon Cancer Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(18): 118-123.
DOI：

YU Wen-yan, WANG Hua-ying, WANG Guo-juan, et al. Effect of Shenling Baizhu San Combined with Oxaliplatin on Proliferation and Apoptosis of Human Colon Cancer Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(18): 118-123. DOI： 10.13422/j.cnki.syfjx.20181716.

摘要

目的：探讨参苓白术散（Shenling Baizhu San，SBS）对人结肠癌HCT116细胞增殖及凋亡的影响。方法：以化疗药奥沙利铂（oxaliplatin，L-OHP）为阳性药，采用噻唑蓝[3-（4，5-dimethyl-2-thiazolyl）-2，5-diphenyl-2H-tetrazolium bromide，MTT]比色法检测SBS联用L-OHP前后对HCT116细胞生长的抑制情况，碘化丙啶（PI）染色分析细胞周期分布，Annexin V-FITC试剂盒检测HCT116细胞凋亡，蛋白免疫印迹法（Western blot）检测其对细胞内B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2），Bcl-2相关X蛋白（Bax）蛋白的影响。结果： ① MTT比色法结果显示，分别取0.002，0.004，0.008，0.016，0.032，0.064，0.128 g·L-1 L-OHP联用5% SBS药物血清作用于HCT116细胞48 h，与联用前各组比较，联用后各组细胞抑制率均明显上升（P<0.05），其中以0.064 g·L-1L-OHP联用5% SBS最高，达（94.95±0.90）%；②周期结果显示，与联用前L-OHP （0.002，0.004，0.008 g·L-1）组比较，分别联用5% SBS后，G2/M期HCT116细胞分布增多，均由G0/G1，S和G2/M期转至G2/M期，以L-OHP 0.008 g·L-1联用5% SBS组最为明显（P<0.05）；③凋亡结果显示，与联用前L-OHP （0.002，0.004，0.008 g·L-1）组比较，分别联用5% SBS后均能明显增加HCT116细胞凋亡率（P<0.05），以高剂量联用组凋亡率最高，为（24.97 ±2.45）%；④蛋白检测显示，与5%空白血清组比较，L-OHP （0.002，0.004，0.008 g·L-1）分别联用5% SBS后，Bax蛋白表达量明显上调（P<0.05），以L-OHP 0.008 g·L-1联用5% SBS组最高。分别与空白血清组、空白组比较，各用药组Bcl-2蛋白表达量均明显降低（P<0.05），其中以L-OHP 0.008 g·L-1联用5% SBS组最低。结论：参苓白术散能协同L-OHP抑制人结肠癌HCT116细胞增殖及凋亡，其作用机制可能是通过促进HCT116细胞增殖、诱导细胞凋亡、改变细胞周期分布、上调Bax蛋白表达及下调Bcl-2蛋白表达来实现的。

Abstract

Objective: To investigate the effect of Shenling Baizhu San (SBS) on the proliferation and apoptosis of human colon cancer HCT116 cells. Method: The growth of HCT116 cell lines treated with SBS was investigated by 3-(4

5-dimethyl-2-thiazolyl)-2

5-diphenyl-2H-tetrazolium bromide(MTT) assay

the cell cycle distribution was analyzed by flow cytometry propidium iodide (PI) staining

and its apoptosis was monitored by Annexin V-FITC kit.B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) were monitored by Westem blot(WB). Result: ① MTT results showed that the inhibition rates of HCT116 cells treated by the groups of 0.002

0.004

0.008

0.016

0.032

0.064

0.128 g·L-1oxaliplatin combined with 5%SBS serum respectively were all significantly increased compared with those uncombined with 5%SBS serum after 48 hours (P<0.05). The inhibition rate of the group of 0.064 g·L-1in L-OHP combined with 5%SBS serum was the highest or (94.95±0.90)%. ② Cell cycle results showed that the distributions of HCT116 cells were all increased in G2/M phase

which were transferred from G0/G1

S and G2/M phases to G2/M phase after the combined administration of low

medium and high-dose oxaliplatin (0.002

0.004

0.008 g·L-1) with 5%SBS serum respectively

and the high-dose oxaliplatin combined 5%SBS serum was the most significant (P<0.05). ③Apoptosis results showed that the apoptosis of HCT116 cells could be induced by low

medium and high-dose oxaliplatin before and after being combined with 5%SBS serum respectively (P<0.05)

and the high-dose combination group showed the highest apoptosis rate or (24.97 ±2.45)%. ④ WB results showed that compared with the 5%blank serum group

the expression of Bax protein was significantly up-regulated after the combined administration of low

medium and high-dose L-OHP with 5%SBS respectively (P<0.05). Compared with blank serum group and blank control group

the expression of Bcl-2 protein in each drug group was significantly decreased (P<0.05)

and the high-dose combination group showed the lowest expression. Conclusion: SBS could inhibit the proliferation and apoptosis of HCT116 cell lines when being combined with oxaliplatin. The mechanism may be achieved by inhibiting the proliferation

inducing the cell apoptosis

changing the cell cycle distribution

up-regulating the Bax protein expression and down-regulating the Bcl-2 protein expression.

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