Effect of Serum Containing Xiaoai Jiedu Decoction on Proliferation and Glycolysis of Human Colon Cancer Cells

SHI Wen-jing; TAN Jia-ni; SHEN Wei-xing; XU Chang-liang; SUN Dong-dong; CHENG Hai-bo

doi:10.13422/j.cnki.syfjx.2017200120

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Effect of Serum Containing Xiaoai Jiedu Decoction on Proliferation and Glycolysis of Human Colon Cancer Cells

更新时间：2024-01-04

- Effect of Serum Containing Xiaoai Jiedu Decoction on Proliferation and Glycolysis of Human Colon Cancer Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 20, Pages: 120-125(2017)
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- DOI：10.13422/j.cnki.syfjx.2017200120
  CLC： R285
- Published：2017
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SHI Wen-jing, TAN Jia-ni, SHEN Wei-xing, et al. Effect of Serum Containing Xiaoai Jiedu Decoction on Proliferation and Glycolysis of Human Colon Cancer Cells[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(20): 120-125.
DOI：

SHI Wen-jing, TAN Jia-ni, SHEN Wei-xing, et al. Effect of Serum Containing Xiaoai Jiedu Decoction on Proliferation and Glycolysis of Human Colon Cancer Cells[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(20): 120-125. DOI： 10.13422/j.cnki.syfjx.2017200120.

摘要

目的：探讨不同浓度消癌解毒方含药血清对人结肠癌细胞增殖及糖酵解过程的影响及其作用机制。方法： 8只新西兰兔随机分为含药血清组（消癌解毒方40 g · kg-1 · d-1）和空白血清组（等体积生理盐水），每组兔子连续灌胃3 d，制备消癌解毒方含药血清和空白血清。5%，10%，15%消癌解毒方含药血清处理人结肠癌HT-29与HCT-116细胞后，采用噻唑蓝比色法测定其对HT-29与HCT-116细胞生长的抑制作用，乳酸试剂盒检测HT-29与HCT-116细胞的乳酸生成量，葡萄糖试剂盒检测HT-29与HCT-116细胞的葡萄糖生成量，蛋白免疫印迹法（Western blot）检测己糖激酶（HK2），丙酮酸脱氢酶激酶（PDK1），乳酸脱氢酶（LDHA）和低氧诱导因子-1α（HIF-1α）蛋白表达。结果：消癌解毒方含药血清能够抑制HT-29与HCT-116细胞增殖能力，15%消癌解毒方含药血清对人结肠癌HT-29与HCT-116细胞的抑制率分别为42.7%，50.2%。Western blot结果显示，与空白组比较，消癌解毒方含药血清可降低HK2，PDK1，LDHA，HIF-1α蛋白相对表达量（P<0.05），且随浓度的增加表达降低。乳酸及葡萄糖试剂盒检测结果显示，与空白组比较，消癌解毒方含药血清可降低HT-29与HCT-116细胞的乳酸及葡萄糖水平（P<0.05），且随浓度的增加乳酸及葡萄糖水平降低。结论：消癌解毒方含药血清可以通过抑制人结肠癌HT-29与HCT-116细胞糖酵解的过程从而抑制肿瘤细胞的增殖，其抑制糖酵解过程的机制可能与降低HIF-1α表达，进而减少糖酵解相关酶HK2，PDK1，LDHA表达有关。

Abstract

Objective: To explore the effects of the containing serum of Xiaoai Jiedu decoction with different concentrations on proliferation and glycolysis of human colon cancer cells and investigate its action mechanism. Method: Eight new zealand rabbits were randomized into the serum containing medicine[40 g · (kg · d)-1 of Xiaoai Jiedu decoction]group and the blank serum group (equivoluminal normal saline).Both groups received intragastric administration for 3 days to prepare serum containing Xiaoai Jiedu decoction and blank serum.Human colon cancer HT-29 and HCT-116 cells were cultured with pharmacological serum of Xiaoai Jiedu decoction (5%

10% and 15%)

and then their effects on the human colon cancer cells' proliferation was examined by methyl thiazolyl tetrazolium assay.The amount of lactic acid produced by HT-29 and HCT-116 cells was measured by lactic acid test kit

and the amount of glucose produced by HT-29 and HCT-116 cells was measured by glucose test kit.The expression levels of hexokinase 2 (HK2)

pyruvate dehydrogenase kinase 1 (PDK1)

lactate dehydrogenase (LDHA)

and Hypoxia inducible factor-1α (HIF-1α) were detected by using Western blot. Result: Serum containing Xiaoai Jiedu decoction exhibited suppressive effects on HT-29 and HCT-116 proliferation.The effect of 15% serum containing Xiaoai Jiedu decoction was the best

with inhibition rate of 42.7% and 50.2% respectively for HT-29 and HCT-116 cells.Western blot analysis indicated that as compared with blank control group

the serum containing Xiaoai Jiedu decoction reduced the expression of HK2

PDK1

LDHA and HIF-1α (P<0.05)

and with the increase of concentration

the expression was decreased.The results of lactic acid and glucose test kit showed that as compared with the blank control group

the serum containing Xiaoai Jiedu decoction can reduce the levels of lactic acid and glucose (P<0.05) and with the increasing of concentration

the levels of lactic acid and glucose were decreased. Conclusion: The serum containing Xiaoai Jiedu decoction can inhibit the proliferation of human colon cancer HT-29 and HCT-116 cells by inhibiting the process of the glycolysis.Inhibitory mechanisms of glycolysis might be associated with reducing the expression of HIF-1α

thereby reducing the expression of glycolytic enzymes HK2

PDK1

and LDHA.

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