Effects of baicalin combined with cisplatin on proliferation and apoptosis of lung adenocarcinoma A549 and H1299 cells
Objective To observe the effects of baicalin combined with cisplatin on the prolifera-tion and apoptosis of lung adenocarcinoma A549 and H1299 cells.Methods Lung adenocarcinoma A549 and H1 299 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 con-trol 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 ex-periment showed that the number of cloned cells of A549 and H1299 cell lines in blank control group,ba-icalein 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 H1299 cell lines in blank control group,ba-icalein group,cisplatin group and combined group were(4.5±1.6)％,(7.3±3.2)％,(21.2±4.6)％,(36.6±3.8)％vs.(3.8±2.2)％,(7.5±1.4)％,(23.5±3.8)％,(35.2±2.9)％(F=18.110,36.258,P＜0.05).The percentage of apoptotic cells in blank control group,baicalein group,cisplatin group and combined group was(5.56±2.73)％,(22.38±3.16)％,(50.78±4.85)％,(66.08±2.96)％vs.(4.78±1.95)％,(23.75±2.56)％,(52.14±3.82)％,(63.56±3.14)％(F=66.565,89.298,P＜0.01).Western blotting showed that the expression levels of pro-apoptotic proteins Caspase-3,c-Caspase-3 and bax in Baicalin group,cisplatin group and combined group were significantly increased as compared with those in the control group,while the expression levels of anti-apoptotic protein bcl-2 were sig-nificantly decreased as compared with the control group.The tumor formation model of nude mice showed that after 4 weeks,the tumor volume of blank control group,baicalein group,cisplatin group and combined group was(625.78±3.62),(365.64±2.58),(220.25±2.24),(122.69±4.86)mm3.The tumor weight of blank control group,baicalein group,cisplatin group and combined group was(0.72±0.14),(0.44± 0.05),(0.22±0.04),(0.16±0.04)g(F=115.480,145.298,P＜0.01).Conclusion The combina-tion of baicalin and cisplatin can inhibit the proliferation and promote apoptosis of lung adenocarcinoma A549 and H1299 cells,and they have a synergistic effect,mainly through the bax/bcl-2/Caspase-3 apoptosis signal pathway,which up-regulates the expression of Caspase-3 and bax protein,and down-regulates the expression of bcl-2 protein.The apoptosis of tumor cells may be induced by ROS sensitizing cisplatin.
WogoninCisplatinAdenocarcinoma of lungProliferationApoptosis
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