人参皂苷Rh_(2)抑制人非小细胞肺癌细胞增殖的机制研究  

Mechanism of ginsenoside Rh_(2) on inhibiting proliferation of human non-small cell lung cancer cells

作　　者：孙小单[1] 王天鸣 李慧[3] 赵沛妍 刘岩[3] 程颖[4] SUN Xiao-dan;WANG Tian-ming;LI Hui;ZHAO Pei-yan;LIU Yan;CHENG Ying(Postdoctoral Research Workstation,Jilin Cancer Hospital,Changchun 130012,China;College of Traditional Chinese Medicine,Changchun University of Chinese Medicine,Changchun 130117,China;Laboratory of Translational Cancer Research,Jilin Cancer Hospital,Changchun 130012,China;Department of Medical Thoracic Oncology,Jilin Cancer Hospital,Changchun 130012,China)

机构地区：[1]吉林省肿瘤医院博士后科研工作站,吉林长春130012 [2]长春中医药大学中医学院,吉林长春130117 [3]吉林省肿瘤医院肿瘤转化医学实验室,吉林长春130012 [4]吉林省肿瘤医院胸部肿瘤内科,吉林长春130012

出　　处：《中草药》2022年第2期441-448,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(82103343);吉林省中医药科技项目(2021095);吉林省卫生计生委科研项目(2018J021);吉林省科技厅项目(202002063JC)。

摘　　要：目的考察人参皂苷Rh_(2)对人非小细胞肺癌A549及H460细胞增殖的影响,并进一步探讨其作用机制。方法人参皂苷Rh_(2)处理A549及H460细胞,采用CCK-8法及克隆形成实验检测药物对细胞增殖的作用;观察药物对细胞形态的影响,利用Annexin V-FITC/碘化丙啶(PI)双染法检测细胞凋亡情况;利用乳酸检测试剂盒检测药物对细胞糖酵解产物即乳酸生成的影响;采用Western blotting法检测药物对线粒体凋亡蛋白[B淋巴细胞瘤2 (B-cell lymphoma 2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、半胱氨酸蛋白酶-3 (Caspase-3)],糖酵解调节通路信号传导与转录激活因子3 (signal transducer and activator of transcription 3,STAT3)/c-myc及糖酵解途径关键酶丙酮酸激酶M2 (pyruvate kinase M2,PKM2)、乳酸脱氢酶A (lactate dehydrogenase A,LDHA)表达的影响。上调c-myc表达后,观察人参皂苷Rh;对A549及H460细胞乳酸水平的影响。结果人参皂苷Rh;可明显抑制A549及H460细胞的增殖能力及克隆形成能力(P<0.05、0.01、0.001),呈剂量和时间相关性。人参皂苷Rh;明显改变A549及H460细胞形态并诱导细胞凋亡(P<0.05),显著降低乳酸生成量(P<0.05、0.01)。人参皂苷Rh;可显著下调Bcl-2/Bax、PKM2、LDHA蛋白表达水平(P<0.05、0.01),上调Caspase-3蛋白表达水平(P<0.05),并抑制STAT3/c-myc通路活性(P<0.05、0.01)。上调c-myc表达后,人参皂苷Rh_(2)对A549及H460细胞乳酸水平的抑制作用减弱(P<0.05、0.01)。结论人参皂苷Rh_(2)显著抑制A549及H460细胞的增殖,其机制可能是通过调控线粒体凋亡蛋白、抑制STAT3/c-myc通路及糖酵解关键酶的表达干扰糖酵解,最终导致细胞凋亡。Objective To study the effect and mechanism of ginsenoside Rh_(2)on proliferation of human non-small cell lung cancer A549 and H460 cells. Methods A549 and H460 cells were treated with ginsenoside Rh_(2), CCK-8 assay and colony formation experiment were used to detect the effect of ginsenoside Rh_(2)on cell proliferation;Effect of ginsenoside Rh_(2)on cell morphology was observed;Cell apoptosis were detected by Annexin V-FITC/PI dye method;Level of glycolytic products(lactic acid) was tested by lactic acid detection kit. Protein expression levels of mitochondrial apoptotic protein B-cell lymphoma 2(Bcl-2), Bcl-2 associated X protein(Bax), Caspase-3 and glycolysis regulation pathway signal transducer and activator of transcription 3(STAT3)/c-myc and glycolysis key enzymes pyruvate kinase M2(PKM2) and lactate dehydrogenase A(LDHA) were detected by Western blotting. Effect of ginsenoside Rh_(2)on lactate level in A549 and H460 cells after up-regulating c-myc expression were observed. Results Ginsenoside Rh_(2)significantly inhibited the proliferation and colony formation ability of A549 and H460 cells in a concentration and time-dependent way(P < 0.05, 0.01, 0.001). Ginsenoside Rh;altered cell morphology and significantly induced cell apoptosis(P < 0.05) and reduced the levels of lactic acid(P < 0.05, 0.01). Ginsenoside Rh_(2)down-regulated Bcl-2/Bax, PKM2 and LDHA protein expressions, upregulated Caspase-3 expression(P < 0.05) and inhibited STAT3/c-myc pathway activity(P < 0.05, 0.01). The inhibitory effect of ginsenoside Rh;on lactate levels in A549 and H460 cells was abolished after up-regulating c-myc expression. Conclusion Ginsenoside Rh_(2)has a significant inhibitory effect on proliferation of A549 and H460 cells. The mechanism may be through regulating mitochondrial apoptotic proteins, inhibiting STAT3/c-myc pathway and reducing glycolysis key enzyme expression to interfere glycolysis, and then leading to cells apoptosis.

关 键 词：人参皂苷Rh_(2) 非小细胞肺癌 增殖 凋亡 糖酵解 信号传导与转录激活因子3/c-myc通路 

分 类 号：R285.5[医药卫生—中药学]