汉黄芩素联合顺铂对肺腺癌A549、H1299细胞增殖和凋亡的影响  

Effects of baicalin combined with cisplatin on proliferation and apoptosis of lung adenocarcinoma A549 and H1299 cells

作　　者：郭晨昊 朱凯丽 鲁明 曹梦悦 苏飞 张涛 侯小明[1,2] 芦永斌[3] 令晓玲 Guo Chenhao;Zhu Kaili;Lu Ming;Cao Mengyue;Su Fei;Zhang Tao;Hou Xiaoming;Lu Yongbin;Ling Xiaoling(The First Clinical Medical College of Lanzhou University,Lanzhou 730000,China;Department of Oncology,the First Hospital of Lanzhou University,Lanzhou 730000,China;Center of Evidence-based Medicine,Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一临床医学院,兰州730000 [2]兰州大学第一医院肿瘤内科,兰州730000 [3]兰州大学循证医学中心,兰州730000

出　　处：《中华实验外科杂志》2024年第4期767-772,共6页Chinese Journal of Experimental Surgery

基　　金：甘肃省重点基地建设项目、甘肃省胸部肿瘤临床研究中心(21JR7RA390);甘肃省科技厅重点研发项目(22YF7FA086);甘肃省科技厅联合科研基金(23JRRA1497);兰州市科技发展指导性项目(2020-ZD-74);兰州市人才创新创业项目(2023-QN-14);兰州大学第一医院院内学科交叉项目(ZX-62000002-2022-532)。

摘　　要：目的观察汉黄芩素联合顺铂对肺腺癌A549、H1299细胞增殖和凋亡的影响。方法培养建立肺腺癌A549、H1299细胞株,实验分为空白对照组、汉黄芩素组、顺铂组及汉黄芩素联合顺铂组(联合组)。采用细胞计数试剂盒(CCK-8)实验和5-乙炔基-2’脱氧尿嘧啶核苷(EdU)实验检测各组的细胞增殖能力;平板克隆实验检测癌细胞克隆形成能力;原位缺口末端标记法(TUNEL)实验检测各组细胞凋亡率;流式细胞术分析各组对细胞凋亡的影响;蛋白质印迹法(Western blot)实验检测B细胞淋巴瘤/白血病-2相关X蛋白(bax)/B细胞淋巴瘤/白血病-2(bcl-2)/半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3通路相关蛋白表达;同时构建SPF级裸鼠成瘤模型进一步验证。多组资料比较采用单因素方差分析。结果CCK-8实验结果显示第4天时肺腺癌A549、H1299细胞株空白对照组、汉黄芩素组、顺铂组、联合组在波长450 nm处时吸光度值比较,差异有统计学意义(1.102±0.109、0.765±0.087、0.484±0.122、0.356±0.056比1.114±0.256、0.824±0.225、0.623±0.239、0.378±0.191,F=259.633、205.262,P<0.01)。EdU实验结果显示肺腺癌A549、H1299细胞株空白对照组、汉黄芩素组、顺铂组、联合组EdU阳性细胞率比较,差异有统计学意义[(58.0±2.0)%、(48.8±3.2)%、(22.5±1.4)%、(16.1±3.8)%比(56.7±1.5)%、(46.6±2.8)%、(21.2±2.2)%、(14.2±1.8)%,F=65.198、89.293,P<0.01]。克隆实验结果显示肺腺癌A549、H1299细胞株空白对照组、汉黄芩素组、顺铂组、联合组克隆细胞数比较,差异有统计学意义(102.0±3.5、96.0±4.8、50.0±2.9、25.0±3.5比125.0±0.8、121.0±2.5、55.0±3.9、30.0±3.3,F=68.151、52.086,P<0.01)。TUNEL实验结果显示肺腺癌A549、H1299细胞株空白对照组、汉黄芩素组、顺铂组、联合组TUNEL阳性细胞率比较,差异有统计学意义[(4.5±1.6)%、(7.3±3.2)%、(21.2±4.6)%、(36.6±3.8)%比(3.8±2.2)%、(7.5±1.4)%、(23.5±3.8)%、(35.2±2.9)%Objective To observe the effects of baicalin combined with cisplatin on the proliferation and apoptosis of lung adenocarcinoma A549 and H1299 cells.Methods Lung adenocarcinoma A549 and H1299 cells were cultured and divided into blank control group,baicalein group,cisplatin group and baicalein+cisplatin group(combined group).The cell proliferation ability of each group was detected by cell counting kit-8(CCK-8)assay and 5-Ethynyl-2′-deoxyuridine(EdU)assay.The cloning ability of cancer cells was detected by plate cloning assay.Terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)assay was used to detect the apoptosis rate of each group.The flow cytometry was used to analyze apoptosis in each group.Western blotting was used to detect the expression of B cell lymphoma/leukemia-2 associated X protein(bax)/B cell lymphoma/leukemia-2(bcl-2)/cysteinyl aspartate-specific protease(Caspase)-3 pathway related proteins.At the same time,SPF nude mouse tumor formation model was established for further verification.Multivariate data were compared by one-way ANOVA.Results The results of CCK-8 assay showed that the absorbance values of A549 and H1299 cells in the blank control group,baicalin group,cisplatin group and combined group were 1.102±0.109,0.765±0.087,0.484±0.122,0.356±0.056 vs.1.114±0.256,0.824±0.225,0.623±0.239,0.378±0.191(F=259.633,205.262,P<0.01).The EdU test results showed that the EdU positive cell rates of A549 and H1299 cells in blank control group,baicalein group,cisplatin group and combined group were(58.0±2.0)%,(48.8±3.2)%,(22.5±1.4)%,(16.1±3.8)%vs.(56.7±1.5)%,(46.6±2.8)%,(21.2±2.2)%,(14.2±1.8)%(F=65.198,89.293,P<0.01).The results of cloning experiment showed that the number of cloned cells of A549 and H1299 cell lines in blank control group,baicalein group,cisplatin group and combined group was 102.0±3.5,96.0±4.8,50.0±2.9,25.0±3.5 vs.125.0±0.8,121.0±2.5,55.0±3.9,30.0±3.3(F=68.151,52.086,P<0.01).TUNEL test results showed that the TUNEL positive cell rates of A549 and

关 键 词：汉黄芩素 顺铂 肺腺癌 增殖 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]