中药组方黄芩汤抑制胃癌细胞增殖的实验研究

doi:10.3877/cma.j.issn.2095-1221.2019.06.004

中华细胞与干细胞杂志(电子版) ›› 2019, Vol. 09 ›› Issue (06) : 340 -344. doi: 10.3877/cma.j.issn.2095-1221.2019.06.004

所属专题：文献；

论著

中药组方黄芩汤抑制胃癌细胞增殖的实验研究

林少琴¹, 张绍琴², 陈思玉¹, 陈舒怡¹, 陈雄³, 欧阳学农³^,^†(

)

^1. 350025　福州，联勤保障部队第九〇〇医院肿瘤科；350122　福州，福建中医药大学药学院
^2. 350003　福州，福建省肿瘤医院中西医结合科
^3. 350025　福州，联勤保障部队第九〇〇医院肿瘤科

收稿日期:2019-10-12 出版日期:2019-12-01
通信作者: 欧阳学农
基金资助:
国家自然科学基金(81274002); 福建省自然科学基金(2016J01586)

Experimental study of traditional Chinese medicine Huangqin Decoction in inhibition of gastric cancer cell proliferation

Shaoqin Lin¹, Shaoqin Zhang², Siyu Chen¹, Shuyi Chen¹, Xiong Chen³, Xuenong Ouyang³^,^†()

^1. Department of Oncology, 900 Hospital of the Joint Logistics Team, Fuzhou 350025, China; College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350025, China
^2. Department of Integrated Traditional Chinese and Western Medicine, Fujian Cancer Hospital, Fuzhou 350025, China
^3. Department of Oncology, 900 Hospital of the Joint Logistics Team, Fuzhou 350025, China

Received:2019-10-12 Published:2019-12-01
Corresponding author: Xuenong Ouyang
About author:
Corresponding author: Ouyang Xuenong, Email:oyxnong@163.com

全文 ( ) 下载PDF ( ) 导出引用

引用本文：

林少琴, 张绍琴, 陈思玉, 陈舒怡, 陈雄, 欧阳学农. 中药组方黄芩汤抑制胃癌细胞增殖的实验研究[J]. 中华细胞与干细胞杂志(电子版), 2019, 09(06): 340-344.

Shaoqin Lin, Shaoqin Zhang, Siyu Chen, Shuyi Chen, Xiong Chen, Xuenong Ouyang. Experimental study of traditional Chinese medicine Huangqin Decoction in inhibition of gastric cancer cell proliferation[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2019, 09(06): 340-344.

目的

探讨黄芩汤对胃癌细胞生长增殖及干细胞标志物表达的影响。

方法

采用体外优化培养的胃癌细胞株SGC-7901模型，分别加入生理盐水、5-FU、黄岑汤和5-FU+黄芪汤干预，MTT法检测胃癌细胞增殖情况，Real-time PCR检测胃癌干细胞相关表面标志物CD44、EpCAM、CD90的表达情况；并进一步利用胃癌细胞裸鼠荷瘤模型，观察黄岑汤协同5-FU对体内肿瘤生长的抑制效果。细胞增殖曲线多组间数据比较采用重复测量数据方差分析，单时间点多组数据采用单因素方差分析，组间比较采用Tukey法。

结果

优化培养的胃癌细胞SGC- 7901呈球形生长，细胞活性染色显示良好的活性状态；MTT结果显示，黄岑汤组（0.44±0.04）能够抑制胃癌细胞的增殖，从第4天起与对照组（0.59±0.02）相比，差异具有统计学意义（F = 39.550，P < 0.01）；黄岑汤联合5-FU组（0.36±0.04）加强5-FU对肿瘤细胞的抑制作用，从第3天起与对照组（0.50±0.01）相比，差异具有统计学意义（F = 10.670，P < 0.01），与5-FU组（0.42±0.03）对比，差异具有统计学意义（F = 10.670，P < 0.05）；Real-time PCR结果显示黄岑汤联合5-FU可抑制胃癌细胞中肿瘤干细胞相关表面标志物CD44、EpCAM、CD90表达（P < 0.01）；移植瘤生长抑制实验发现黄芪汤组对荷瘤鼠移植瘤的生长具有抑制作用，与对照组相比移植瘤体积减小，差异具有统计学意义（P < 0.01），黄岑汤联合5-FU组与5-FU组相比对移植瘤抑制效果更明显，差异具有统计学意义（P < 0.01）。

结论

黄芪汤能抑制胃癌细胞的生长和增殖，并能协同5-FU产生抗肿瘤作用。

关键词: 胃癌细胞, 黄芩汤, 细胞增殖, 5-氟尿嘧啶

Objective

To investigate the effect of Huangqin Decoction on the growth and proliferation of gastric cancer cells and the expression of stem cell markers.

Methods

The gastric cancer cell line SGC-7901, which was optimally cultivated in vitro, was added with physiological saline, 5-FU, Huangqin Decoction, and 5-FU + Huangqin Decoction for intervention. The proliferation of gastric cancer cells was detected by MTT assay, and the surface markers associated with gastric cancer stem cells, CD44, EpCAM and CD90, were detected by Real-time PCR. Furthermore, a nude mouse model bearing gastric cancer cells was used to observe the inhibitory effect of Huangqin Decoction in combination with 5-FU on tumor growth in vivo. Cell proliferation data were analyzed by repeated measures multivariate analysis of variance, data comparison between multiple groups was made by using one-way ANOVA, and comparison between groups by using Tukey's method.

Results

The optimally cultured gastric cancer cells SGC-7901 showed a spherical growth, and the cell viability staining showed a good active state. The MTT results showed that the Huangqin Decoction group (0.44±0.04) could significantly inhibit the proliferation of gastric cancer cells, compared with the control group (0.59±0.02) from the fourth day There was a statistical difference (P < 0.01) ; Huangqin Decoction combined with 5-FU group (0.36±0.04) could significantly enhance the inhibitory effect of 5-FU on tumor cells, and there was a statistical difference from the control group (0.50±0.01) (P < 0.01) and the 5-FU group (0.42±0.03) on day 3 (P < 0.05) ; Real-time PCR results showed that Huangqin Decoction combined with 5-FU could inhibit tumor stem cell-related surface markers CD44, EpCAM, and CD90 in gastric cancer cells. Expression (P < 0.01) ; the tumor growth inhibition experiment found that the Huangqin Decoction group had a significant inhibitory effect on the growth of tumor-bearing mice, and the tumor volume was decreased significantly compared with the control group (P < 0.01) , Huangqin Decoction combined with 5-FU has a more significant inhibitory effect on transplanted tumors compared with the 5-FU group (P < 0.01) .

Conclusion

Huangqin Decoction can inhibit the growth and proliferation of gastric cancer cells, and exert an antitumor effect when combined with 5-FU.

Key words: Gastric cancer cells, Huangqin Decoction, Cell proliferation, 5-Fluorouracil

表1 实时荧光定量PCR引物序列

基因	Forward primer	Reverse primer
CD44	GCGTAACGCGTCCCGGACACCA	GCGAGGCTTTCAACACAACC
EpCAM	CGCAGCTCAGGAAGAATGTG	TGAAGTACACTGGCATTGACGA
CD90	CCGACTTCCGAAGCACACAGT	GATGCACAAGGAGTGCGGGATCC
GAPDH	CAAGGTCATCCATGACAACTTTG	GTCCACCACCCTGTTGCTGTAG

表1 实时荧光定量PCR引物序列

基因	Forward primer	Reverse primer
CD44	GCGTAACGCGTCCCGGACACCA	GCGAGGCTTTCAACACAACC
EpCAM	CGCAGCTCAGGAAGAATGTG	TGAAGTACACTGGCATTGACGA
CD90	CCGACTTCCGAAGCACACAGT	GATGCACAAGGAGTGCGGGATCC
GAPDH	CAAGGTCATCCATGACAACTTTG	GTCCACCACCCTGTTGCTGTAG

图1 倒置荧光显微镜下观察不同培养时间点胃癌细胞形态变化（FDA/PI双荧光染色，×100）

表2 黄芩汤抑制胃癌细胞增殖情况比较（±s，n = 3）

分组	第1天	第2天	第3天	第4天	第5天	第6天	第7天
对照组	0.41±0.01	0.43±0.02	0.50±0.01	0.59±0.02	0.74±0.03	0.79±0.02	0.82±0.04
黄岑汤组	0.41±0.04	0.41±0.02	0.45±0.03	0.44±0.04^a	0.45±0.04^a	0.53±0.01^a	0.54±0.03^a
5-FU组	0.41±0.03	0.40±0.01	0.42±0.03^a	0.44±0.03^a	0.49±0.03^a	0.49±0.02^a	0.50±0.03^a
黄岑汤+5-FU组	0.41±0.04	0.40±0.03	0.36±0.04^a	0.33±0.02^ab	0.30±0.04^ab	0.29±0.03^ab	0.28±0.03^ab
F值	0.030	0.992	10.670	39.550	75.940	253.200	128.400
P值	0.992	0.444	< 0.01	< 0.01	< 0.01	< 0.01	< 0.01

表2 黄芩汤抑制胃癌细胞增殖情况比较（±s，n = 3）

分组	第1天	第2天	第3天	第4天	第5天	第6天	第7天
对照组	0.41±0.01	0.43±0.02	0.50±0.01	0.59±0.02	0.74±0.03	0.79±0.02	0.82±0.04
黄岑汤组	0.41±0.04	0.41±0.02	0.45±0.03	0.44±0.04^a	0.45±0.04^a	0.53±0.01^a	0.54±0.03^a
5-FU组	0.41±0.03	0.40±0.01	0.42±0.03^a	0.44±0.03^a	0.49±0.03^a	0.49±0.02^a	0.50±0.03^a
黄岑汤+5-FU组	0.41±0.04	0.40±0.03	0.36±0.04^a	0.33±0.02^ab	0.30±0.04^ab	0.29±0.03^ab	0.28±0.03^ab
F值	0.030	0.992	10.670	39.550	75.940	253.200	128.400
P值	0.992	0.444	< 0.01	< 0.01	< 0.01	< 0.01	< 0.01

表3 黄岑汤抑制胃癌细胞肿瘤干细胞表面标志物表达（±s，n = 3）

分组	CD44	EpCAM	CD90
对照组	1.00±0.02	1.00±0.03	1.00±0.02
黄岑汤组	0.95±0.02	0.80±0.03^a	0.85±0.02^a
5-FU组	0.86±0.03^a	0.74±0.02^a	0.82±0.03^a
黄岑汤+5FU组	0.45±0.03^ab	0.36±0.02^ab	0.43±0.02^ab
F值	276.300	315.000	412.200
P值	< 0.01	< 0.01	< 0.01

表3 黄岑汤抑制胃癌细胞肿瘤干细胞表面标志物表达（±s，n = 3）

分组	CD44	EpCAM	CD90
对照组	1.00±0.02	1.00±0.03	1.00±0.02
黄岑汤组	0.95±0.02	0.80±0.03^a	0.85±0.02^a
5-FU组	0.86±0.03^a	0.74±0.02^a	0.82±0.03^a
黄岑汤+5FU组	0.45±0.03^ab	0.36±0.02^ab	0.43±0.02^ab
F值	276.300	315.000	412.200
P值	< 0.01	< 0.01	< 0.01

表4 各组荷瘤鼠移植瘤的体积（mm³，±s）

分组	动物数（只）	第0天	第15天
对照组	5	149.52±63.97	506.73±17.26
5-FU组	5	142.68±35.72	380.70±24.63^a
黄芪汤组	5	144.70±41.06	407.33±18.29^a
5-FU+黄芪汤组	5	146.29±28.82	301.84±19.74^ab
F值	?	0.008	92.250
P值	?	0.998	< 0.01

表4 各组荷瘤鼠移植瘤的体积（mm³，±s）

分组	动物数（只）	第0天	第15天
对照组	5	149.52±63.97	506.73±17.26
5-FU组	5	142.68±35.72	380.70±24.63^a
黄芪汤组	5	144.70±41.06	407.33±18.29^a
5-FU+黄芪汤组	5	146.29±28.82	301.84±19.74^ab
F值	?	0.008	92.250
P值	?	0.998	< 0.01

图2 荷瘤裸鼠体内移植瘤大小

1	Rawla P, Barsouk A. Epidemiology of gastric cancer: global trends, risk factors and prevention[J]. Prz Gastroenterol, 2019, 14(1):26-38.
2	Wang C, Tang X, Zhang L. Huangqin-Tang and Ingredients in Modulating the Pathogenesis of Ulcerative Colitis[J]. Evid Based Complement Alternat Med, 2017, 2017:7016468.
3	Zou Y, Li WY, Wan Z, et al. Huangqin-Tang Ameliorates TNBS-Induced Colitis by Regulating Effector and Regulatory CD4(+) T Cells [J]. Biomed Res Int, 2015, 2015:102021.
4	Tilton R, Paiva AA, Guan JQ, et al. A comprehensive platform for quality control of botanical drugs (PhytomicsQC):a case study of Huangqin Tang (HQT) and PHY906[J]. Chin Med, 2010, 5:30.
5	Loh TJ, Moon H, Jang HN, et al. SR proteins regulate V6 exon splicing of CD44 pre-Mrna [J]. BMB Rep, 2016, 49(11):612-616.
6	Bekaii-Saab T, El-Rayes B. Identifying and targeting cancer stem cells in the treatment of gastric cancer[J]. Cancer 2017,123(8):1303-1312.
7	Petrovchich I, Ford JM. Genetic predisposition to gastric cancer[J]. Semin Oncol, 2016, 43(5):554-559.
8	Obermannová R, Lordick F. Management of metastatic gastric cancer [J]. Hematol Oncol Clin North Am, 2017, 31(3):469-483.
9	Wang DF, Wang YL, Wang YW, et al. Effect of Huangqin Tang on the function of regulatory TLR4/MyD88 signal pathway in rats with ulcerative colitis[J]. Yao Xue Xue Bao, 2016, 51(10):1558-1563.
10	Wang YW, Zhang HH, Wang YL, et al. Effect of huangqin tang on the regulatory NF-κB p65 signal pathway in rats with ulcerative colitis[J]. Yao Xue Xue Bao, 2015, 50(1):21-27.
11	Bonham M, Posakony J, Coleman I, et al. Characterization of chemical constituents in Scutellaria baicalensis with antiandrogenic and growth-inhibitory activities toward prostate carcinoma[J]. Clin Cancer Res, 2005, 11(10):3905-3914.
12	Sancho P, Barneda D, Heeschen C. Hallmarks of cancer stem cell metabolism[J]. Br J Cancer, 2016, 114(12):1305-1312.

[1]	程慧, 李妍雨, 张蓓, 成杰, 张艳玲. 微小RNA-195靶向趋化因子5抑制滋养细胞增殖、迁移和侵袭及其机制研究[J]. 中华妇幼临床医学杂志(电子版), 2022, 18(02): 165-174.
[2]	孙佳辰, 宋垚垚, 申传安, 赵虹晴, 孙天骏. 表皮和表皮干细胞衰老的研究进展[J]. 中华损伤与修复杂志(电子版), 2022, 17(06): 531-534.
[3]	江振剑, 蒋明, 黄大莉. TK1、Ki67蛋白在分化型甲状腺癌组织中的表达及预后价值研究[J]. 中华普外科手术学杂志(电子版), 2023, 17(06): 623-626.
[4]	石浩伟, 郝少龙, 纪宇, 孙浩, 聂芳, 胡阳, 李泽乾, 韩威. 长链非编码RNA-BANCR在胰腺癌中的表达及临床意义[J]. 中华普外科手术学杂志(电子版), 2022, 16(05): 554-559.
[5]	雷震, 郭正辉, 唐晨, 彭圣萌, 任艳婷, 吴宛桦, 周杰, 陈勇明, 李凌峰, 黄海, 赖义明. ASF1B通过调控P53相关信号通路促进前列腺癌迁移和增殖的研究[J]. 中华腔镜泌尿外科杂志(电子版), 2022, 16(03): 262-269.
[6]	李芬, 黄文娟, 朱乐攀. 色素上皮因子表达对肺癌细胞增殖及迁移能力的影响[J]. 中华肺部疾病杂志(电子版), 2022, 15(02): 246-248.
[7]	刘燕, 叶亚萍, 郑艳莉. 干扰LINC00466通过miR-493-3p/MIF抑制子宫内膜癌RL95-2细胞恶性生物学行为[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(03): 151-158.
[8]	梁芳, 刘广申, 徐艳. LncRNA AC130710通过miR-129-5P/WNT4轴促进子宫内膜癌细胞增殖和上皮间质转化[J]. 中华细胞与干细胞杂志(电子版), 2022, 12(04): 206-214.
[9]	董会月, 张晓, 祝玲, 张怡, 孙晶晶, 路君. TET3促进肾透明细胞癌细胞增殖[J]. 中华细胞与干细胞杂志(电子版), 2021, 11(06): 337-342.
[10]	莫钊鸿, 翟航, 苏日顺, 孟泓宇, 罗豪, 陈文豪, 许瑞云. U2AF2表达对肝细胞癌增殖和迁移的影响及其与预后的关系[J]. 中华肝脏外科手术学电子杂志, 2023, 12(03): 336-341.
[11]	魏志鸿, 郭娟, 江哲龙, 江艺, 吕立志. miR-4458靶向结合BZW2对肝癌细胞增殖、迁移和侵袭的影响[J]. 中华肝脏外科手术学电子杂志, 2023, 12(01): 108-113.
[12]	汤永昌, 袁峰, 梁豪, 钟昭众, 熊志勇, 曹明波, 任昱朋, 李宇轩, 姚志成, 邓美海. HBx对HBV相关性肝癌增殖和迁移能力的影响及其机制[J]. 中华肝脏外科手术学电子杂志, 2022, 11(02): 198-202.
[13]	杨翠萍, 杨晓金, 全旭, 谢玲, 吴云林, 陈平. 肝细胞核因子-1α基因突变协同腺瘤样结肠息肉病基因突变对家族性腺瘤性息肉病细胞增殖的影响[J]. 中华消化病与影像杂志(电子版), 2022, 12(04): 228-231.
[14]	张懿炜, 胡亚欣, 出良钊, 严昭, 曾茜, 蒲茜. CREB3通过下调FAK磷酸化水平抑制胶质瘤细胞增殖及侵袭转移的体外实验研究[J]. 中华临床医师杂志(电子版), 2023, 17(02): 202-209.
[15]	郭德华, 贺迎坤, 白卫星, 何艳艳, 李天晓. 脑动静脉畸形部分栓塞术后血管组织增殖与凋亡的变化[J]. 中华介入放射学电子杂志, 2022, 10(02): 152-157.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Experimental study of traditional Chinese medicine Huangqin Decoction in inhibition of gastric cancer cell proliferation

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Experimental study of traditional Chinese medicine Huangqin Decoction in inhibition of gastric cancer cell proliferation