消癌平联合热疗对吉非替尼耐药肺腺癌细胞株A549作用的实验研究

The effect of Xiaoaiping combined Thermochemistry on the gefitinib-resistant lung adenocarcinoma cell line A549:a experimental study

目的观察消癌平联合热疗作用于耐吉非替尼人肺腺癌细胞株A549的影响及其作用机制,为临床上消癌平联合热疗治疗非小细胞肺癌提供理论依据。方法以体外培养的耐吉非替尼人肺腺癌细胞株A549,分别将96孔板置于:15%CO2,37℃培养;2加入消癌平药物;35%CO2,42℃培养;45%CO2,42℃联合消癌平进行培养,用MTT法检测A549细胞增殖抑制作用;用RT-PCR方法检测A549细胞中VEGF、b FGF基因表达;用ELISA法检测A549细胞中VEGF、b FGF蛋白含量。结果消癌平单药抑制率为12%,高温组抑制率为12%,消癌平联合高温组抑制率为45%,差异有统计学意义(P<0.05),显示消癌平联合高温组较消癌平、高温组增殖抑制作用增强;消癌平单药组VEGF表达为78.95%,高温组42℃为37.02%,消癌平联合高温组为24.82%,差异有统计学意义(P<0.05);消癌平单药组VEGF蛋白含量表达为60.87%,高温42℃为4.78%,消癌平联合高温组为0.35%,差异有统计学意义(P<0.05);消癌平单药组b FGF蛋白含量表达为95.01%,高温组为97.50%,消癌平联合高温组为78.37%,差异无统计学意义(P>0.05),显示消癌平联合高温组较消癌平、高温组的VEGF、b FGF表达均有下降,且消癌平联合热疗对VEGF蛋白表达抑制作用更强。结论消癌平联合热疗可以逆转吉非替尼NSCLC耐药,其作用机制可能是降低VEGF蛋白表达介导肿瘤血管生成,从而抑制肿瘤细胞增殖。

Objective To observe the effects and mechanisms of Xiaoaiping combined with thermotherapy on gefitinib-re- sistance lung adenocarcinoma cell line A549, to provide theory evidence in thermochemotherapy for clinic. Methods The cultivated gefitinib-resistance lung adenocarcinoma cell line A549 was treated with ①5% CO2 ,37 ℃ training;②to join xiaoaiping;③5% CO2 ,42 ℃ cuhure;④5% CO2,42 ℃ joint xiaoaiping. MTT assay was applied to analyze the inhibiting effects on the proliferation of A549 cells; RT-PCR, ELISA was performed to detected the content of VEGF, bFGF. Results The inhibition rate is 12% for Xiaoaiping,the inhibition rate is 12% for temperature 42 ℃ ,the inhibition rate is 45% for Xiaoaiping combined thermochemistry group, significant differences were observed （ P 〈 0.05 ）, so Xiaoaiping combined thermochemistry group have higher inhibiting effects on the proliferation compared Xiaoaiping, Thermochemistry group ; the expression of VEGF is 78.95 % for Xiaoaiping, the inhibition rate is 37.02% for temperature 42 ℃, the inhibi- tion rate is 24.82% for Xiaoaiping combined thermochemistry group, significant differences were observed（ P 〈 0.05 ） ; The protein expression of VEGF was 60.87% ,4.78%, and 0.35%, respectively, significant differences were observed （P 〈 0.05 ）, while the bFGF has no significant difference. There was an decreased expression of VEGF, bFGF and stron- ger inhibition of VEGF protein expression in Xiaoaiping combined thermochemistry group compared with Xiaoaiping,Ther- mochemistry group. Conclusion Xiaoaiping combined thermochemistry could reverse NSCLC resistance by reducing VEGF protein expression mediated tumor angiogenesis, thus inhibit tumor cell proliferation.