Page 80 - 2020年1月第31卷第2期

二甲双胍对胰腺癌BxPC-3细胞恶性表型的影响 Δ

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黄志铨 ，王振文，朱 亮（南昌大学第一附属医院消化内科，南昌 330006）
#
中图分类号 R965 文献标志码 A 文章编号 1001-0408（2020）02-0202-06
DOI 10.6039/j.issn.1001-0408.2020.02.14

摘 要 目的：探索二甲双胍对胰腺癌BxPC-3细胞恶性表型的影响。方法：以Smad4基因天然缺失型人胰腺癌BxPC-3细胞为对
象，采用CCK-8法和流式细胞术分别检测不同剂量二甲双胍（5、10、20 mmol/L）作用24 h后的细胞增殖和凋亡情况，并计算细胞存
活率和凋亡率；采用Transwell迁移试验检测不同剂量二甲双胍（10、20 mmol/L）作用24 h后的细胞迁移情况，记录迁移细胞数；采
用实时定量聚合酶链反应法和Western blotting法分别检测细胞中钙黏着蛋白E（E-cadherin）、波形蛋白（Vimentin）、补体反应基因
32（RGC-32）的mRNA及蛋白表达情况。结果：与对照组和5 mmol/L二甲双胍组比较，10、20 mmol/L二甲双胍组细胞的存活率均
显著降低，凋亡率均显著升高，且 20 mmol/L 二甲双胍组细胞的凋亡率显著高于 10 mmol/L 二甲双胍组（P＜0.05）。与对照组比
较，10、20 mmol/L 二甲双胍组迁移细胞数均显著减少，且 20 mmol/L 二甲双胍组显著少于 10 mmol/L 二甲双胍组（P＜0.05）；10、
20 mmol/L二甲双胍组细胞中E-cadherin mRNA及其蛋白的相对表达量均显著升高，且20 mmol/L二甲双胍组E-cadherin mRNA
的相对表达量显著高于10 mmol/L二甲双胍组；10 mmol/L二甲双胍组细胞中Vimentin mRNA，20 mmol/L二甲双胍组细胞中Vi-
mentin mRNA 及其蛋白以及 10、20 mmol/L二甲双胍组细胞中 RGC-32 mRNA 及其蛋白的相对表达量均显著降低，且 20 mmol/L
二甲双胍组Vimentin mRNA及其蛋白、RGC-32 mRNA的相对表达量均显著低于10 mmol/L二甲双胍组（P＜0.05或P＜0.01）。结
论：二甲双胍可剂量依赖性地通过Smad4非依赖性通路抑制BxPC-3细胞的增殖和迁移，促进其凋亡，这可能与胰腺癌细胞上皮间
质转化过程以及RGC-32表达受到抑制有关。
关键词 二甲双胍；胰腺癌；BxPC-3细胞；增殖；凋亡；上皮间质转化；补体反应基因32

Effects of Metformin on the Malignant Phenotype of Pancreatic Cancer BxPC-3 Cells
HUANG Zhiquan，WANG Zhenwen，ZHU Liang（Dept. of Gastroenterology，the First Affiliated Hospital of
Nanchang University，Nanchang 330006，China）

ABSTRACT OBJECTIVE：To investigate the effects of metformin on malignant phenotype of pancreatic cancer BxPC-3 cells.
METHODS：Using human pancreatic cancer BxPC-3 cells with natural deletion of Smad4 gene as reaserch objects，CCK-8 assay
and flow cytometry were used to detect the proliferation and apoptosis of BxPC-3 cells after treated with different doses of
metformin（5，10，20 mmol/L）for 24 h. The cell survival rate and apoptosis rate were calculated. Transwell assay was used to test
the migration of cells after treated with different doses of metformin（10，20 mmol/L）for 24 h. The number of migrating cells was
recorded. qRT-PCR and Western blotting assay were performed to determine mRNA and protein expression of E-cadherin，Vimentin
and RGC-32 in cells. RESULTS：Compared with control group and 5 mmol/L metformin group，survival rate of cells were
decreased significantly in 10，20 mmol/L metformin groups，while apoptosis rate was increased significantly；the apoptosis rate in
20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group（P＜0.05）. Compared with control group，
the number of migrating cells was decreased significantly in 10，20 mmol/L metformin groups，and the 20 mmol/L metformin
group was significantly lower than 10 mmol/L metformin group（P＜0.05）. Relative mRNA and protein expression of E-cadherin
were increased significantly in 10，20 mmol/L metformin groups，and relative mRNA expression of E-cadherin in 20 mmol/L
metformin group was significantly higher than 10 mmol/L metformin group. Relative mRNA expression of Vimentin in 10 mmol/L
metformin group，relative mRNA and protein expression of Vimentin in 20 mmol/L metformin group，relative mRNA and protein
expression of RGC-32 in 10，20 mmol/L metformin groups were decreased significantly；relative mRNA and protein expression of
Vimentin as well as mRNA expression of RGC-32 in 20 mmol/L metformin group were significantly lower than 10 mmol/L
metformin group （P＜0.05 or P＜0.01）. CONCLUSIONS：Metformin can inhibit the proliferation and migration of pancreatic
cancer cells through smael-independent pathways in a dose-
Δ 基金项目：江西省青年科学基金资助项目（No.20171BAB215044）；
dependent manner， and promote their apoptosis， which is
江西省教育厅科学技术研究项目（No.170006）；江西省卫生计生委科
associated with the inhibition epithelial-mesenchymal transition
技计划项目（No.20195082）
＊硕士研究生。研究方向：胰腺癌的侵袭和转移机制。电话： and the expression of RGC-32 of pancreatic cancer.
0791-88694228。E-mail：hzq4010@163.com KEYWORDS Metformin；Pancreatic cancer；BxPC-3 cell；
# 通信作者：副主任医师，硕士生导师，博士。研究方向：胰腺癌 Proliferation； Apoptosis； Epithelial-mesenchymal transition；
的侵袭和转移机制。电话：0791-88694228。E-mail：89493075@qq. RGC-32
com

·202 · China Pharmacy 2020 Vol. 31 No. 2 中国药房 2020年第31卷第2期