不同体质血清影响A549肺癌细胞增殖的机理  

Effect of Different Constitutional Types Serum on Mechanism of A549 Lung Cancer Cell Proliferation

作　　者：郑里翔[1] 刘红宁[1] 王萍[1] 付建江[1] 邓佳卉[1] 徐敏娟[1] 王月[1] 

机构地区：[1]江西中医学院,江西南昌330004

出　　处：《时珍国医国药》2012年第6期1523-1524,共2页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.30860348)

摘　　要：目的探讨阴虚、阳虚和平和质3种体质血清对A549肺癌细胞生长的影响,并探讨其部分机理。方法 A549肺癌细胞经平和、阴虚、阳虚体质血清培养24,48,72 h,用MTT、流式细胞的方法分别测定平和、阴虚、阳虚体质血清对A549细胞增殖、细胞周期的变化,并用RT-PCR的方法对周期蛋白D1(CyclinD1)、细胞周期蛋白依赖性蛋白激酶4(CDK4)及P16基因表达的mRNA量的分析。结果细胞培养72 h,阴虚、阳虚体质血清中的A549细胞MTT吸光度值(A)明显高于平和质;流式细胞的结果表明阴虚、阳虚体质血清培养的A549细胞的S期细胞明显高于平和质(P<0.05),而细胞周期G0/G1期的细胞低于平和质((P<0.05);RT-PCR的结果显示,CyclinD1和CDK4基因表达mRNA量均明显高于平和质((P<0.05),而P16基因表达产物均低于平和质((P<0.05)。结论阴虚、阳虚体质血清均可促进A549细胞增殖,可能的机理是提高CyclinD1、Cdk4的表达,降低P16的表达,促进细胞周期从G0/G1向S推进,从而促进细胞增殖。Objective To investigate the effect of gentleness type,＂yin＂ deficiency,＂yang＂ - deficiency serum on A549 cell growth mechanism. Methods Cell culture technique was used in vitro and treated with 10% serum of constitution of gentleness type, yin deficiency, ＂yang＂ - deficiency for 72 h. Cell proliferation was observed by MTT metabolism measuring. Flow cytometry was performed to track cell cycle progression. Cyclin gene expression products were performed to evaluate by RT - PCR. Results The results of MTF assay showed that serum of constitution of ＂yin＂ deficiency, ＂yang＂ - deficiency significantly sitimu- lated cell proliferation than serum of gentleness type did （ P 〈 0.05 ） . Flow cytometry （ FCM ） analysis indicated that the G0/G1 phase fraction ratio of yin deficiency, ＂yang＂ - deficiency serum group was lower than that of serum of gentleness type, while the S - phase fraction ratio was higher than that of serum of gentleness type（ P 〈 0.05 ）. RT - PCR results displayed that serum of con- stitution of ＂yin＂ deficiency,＂yang＂ -deficiency elevated the expression of cyclinDlmRNA,Cdk4mRNA, and decreased the ex- pression of P16 mRNA than serum of gentleness type did（ P 〈 0.05 ）. Conclusion These results showed that serum of constitution of ＂yin＂ deficiency,＂ yang＂ -deficiency significantly sitimulated cell proliferation than serum of gentleness type did. The mecha- nism may be increasing the CyclinD1/Cdk4 gene expression and decreasing P16 gene expression, inducing A549 cell cycle in G0/ G1 phase to S phase transition.

关 键 词：体质血清 癌细胞增殖 细胞周期 周期蛋白基因表达产物 

分 类 号：R285.5[医药卫生—中药学]