粒细胞巨噬细胞集落刺激因子抑制氟尿嘧啶诱导的胃癌细胞凋亡增强耐药性的实验研究

doi:10.3877/cma.j.issn.1674-0793.2020.01.004

中华普通外科学文献(电子版) ›› 2020, Vol. 14 ›› Issue (01) : 10 -13. doi: 10.3877/cma.j.issn.1674-0793.2020.01.004

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论著

粒细胞巨噬细胞集落刺激因子抑制氟尿嘧啶诱导的胃癌细胞凋亡增强耐药性的实验研究

常仕林¹, 何进鹏², 陈亚琼³, 蔡辉⁴^,^†(

)

^1. 750004　银川，宁夏医科大学
^2. 730013　兰州，中国科学院兰州分院空间辐射生物学实验室
^3. 730013　兰州，甘肃省人民医院医务处
^4. 730013　兰州，甘肃省人民医院普外科

收稿日期:2019-08-19 出版日期:2020-02-01
通信作者: 蔡辉

Granulocyte macrophage colony stimulating factor enhancing drug resistance of gastric cancer cells by inhibiting apoptosis

Shilin Chang¹, Jinpeng He², Yaqiong Chen³, Hui Cai⁴^,^†()

^1. Ningxia Medical University, Yinchuan 750004, China
^2. Laboratory of Space Radiation Biology, Lanzhou Branch of Chinese Academy ofSciences, Lanzhou 730013, China
^3. Medical Office, Gansu Provincial People’s Hospital, Lanzhou 730013, China
^4. Department of General Surgery, Gansu Provincial People’s Hospital, Lanzhou 730013, China

Received:2019-08-19 Published:2020-02-01
Corresponding author: Hui Cai
About author:
Corresponding author: Cai Hui, Email: caialonteam@163.com

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目的

通过检测粒细胞巨噬细胞集落刺激因子（GM-CSF）对氟尿嘧啶（5-FU）诱导后胃癌细胞凋亡的影响，初步探讨GM-CSF参与胃癌细胞耐药性的相关机制。

方法

体外培养胃癌SGC7901细胞，MTT法检测5-FU作用于胃癌细胞的半致死剂量；将GM-CSF添加到5-FU处理的胃癌细胞中，MTT法和平板克隆实验检测其对细胞活性和克隆增殖的影响，Annexin V-FITC/PI双染法流式细胞术检测细胞凋亡情况。Western blotting检测凋亡相关蛋白Bax和Bcl-2的表达差异。

结果

5-FU对胃癌细胞的半致死剂量为（16±0.4）mg/L。5-FU对胃癌细胞的活性和增殖能力有明显的抑制作用，GM-CSF能有效减弱5-FU的抑制效果（P<0.05）。5-FU能显著诱导胃癌细胞凋亡，Bax/Bcl-2比值显著升高，而加入GM-CSF后，5-FU诱导的凋亡被抵抗，Bax/Bcl-2比值升高趋势被抑制（P<0.05）。

结论

GM-CSF能够促进胃癌细胞增殖并有效抑制5-FU诱导的胃癌细胞凋亡，在增强胃癌细胞化疗耐药性方面具有重要作用。

关键词: 粒细胞巨噬细胞集落刺激因子, 胃肿瘤, 细胞凋亡, 抗药性,肿瘤, 氟尿嘧啶

Objective

By detecting the effects of granulocyte macrophage colony stimulating factor (GM-CSF) on the apoptosis of gastric cancer cells induced by fluorouracil (5-FU), the mechanism of GM-CSFinvolved in drug resistance of gastric cancer cells was preliminarily explored.

Methods

The gastric cancer SGC7901 cells were cultured in vitro, and the semi-lethal dose of 5-FU on gastric cancer cells was detected by MTT assay. GM-CSF was added to 5-FU-treated gastric cancer cells. The effects of cell proliferation and clonal proliferation were detected by MTT and plate cloning assay. Apoptosis was detected by Annexin V-FITC/PI flow cytometry. Western blotting was used to detect the expression differences of apoptosis-related proteins Bax and Bcl-2.

Results

The semi-lethal dose of 5-FU to gastric cancer cells was (16±0.4) mg/L.5-FU could inhibit the activity and proliferation of gastric cancer cells, which could be effectively weakened by GM-CSF (P<0.05). 5-FU induced apoptosis of gastric cancer cells significantly, Bax/Bcl-2 ratio was significantly increased, while GM-CSF could reverse these trend (P<0.05).

Conclusion

GM-CSF can promote the proliferation of gastric cancer cells and effectively inhibit the apoptosis of gastric cancer cells induced by 5-FU, and plays an important role in enhancing the chemoresistance of gastric cancer cells.

Key words: Granulocyte-macrophage colony-stimulating factor, Stomach neoplasms, Apoptosis, Drug resistance, neoplasm, Fluorouracil

图2 不同处理方式对胃癌SGC7901细胞活性和细胞增殖的影响　A为MTT法检测各组细胞活性率结果，与对照组相比，*P<0.05；联合组与5-FU诱导组相比，*P<0.05。B为克隆形成率，与对照组相比，*P<0.05，**P<0.01；联合组与5-FU诱导组相比，*P<0.05。C为平板克隆实验结果图片；a为空白对照组；b为GM-CSF刺激组；c为5-FU诱导组；d为联合组

图3 倒置相差显微镜下观察四组细胞凋亡情况

图4 不同处理方式对胃癌SGC7901细胞凋亡影响的结果　A为Annexin-V检测细胞凋亡结果；B为四组细胞凋亡率比较，与对照组比较，*P<0.05；联合组与5-FU诱导组相比，*P<0.05

图5 不同处理方式对胃癌SGC7901细胞Bax、Bcl-2蛋白表达的影响　A为各组Bax、Bcl-2蛋白表达结果；B为各组Bax/Bcl-2比值结果；与对照组相比，*P<0.05；联合组与5-FU诱导组相比，*P<0.05

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