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目的观察厄洛替尼联合放射线对MKN45细胞的影响,初步探讨厄洛替尼对其放射增敏机制。方法通过MTT法检测厄洛替尼、5-氟尿嘧啶(5-Fu)对MKN45细胞的生长抑制作用,流式细胞仪检测MKN45细胞经厄洛替尼、5-Fu及联合放射线处理后细胞的凋亡率及周期分布情况;Western Blots法检测厄洛替尼、5-Fu及联合放射线对MKN45细胞的Bax与Bcl-2蛋白表达的影响。结果厄洛替尼、5-Fu均能抑制MKN45细胞的生长,与药物浓度及作用时间呈正相关(P<0.01)。厄洛替尼联合组使G2/M、G0/G1期细胞比率增加最明显,与5-Fu联合组比较有统计学意义(70.32±0.65vs58.25±0.42,P<0.01);细胞凋亡率增加最多,与5-Fu联合组比较有统计学意义(19.21±1.23vs13.11±0.76,P<0.01);厄洛替尼联合组作用于细胞后,Bcl-2蛋白表达明显减少,Bax蛋白表达则明显增加,与其他组比较差异显著(P<0.01)。结论厄洛替尼的放射增敏机制与增加MKN45细胞的细胞凋亡率、G2/M和G0/G1期细胞比率;降低Bcl-2、升高Bax蛋白表达,从而降低Bcl-2/Bax比率有关。
Objective To observe the effect of erlotinib combined with radiation on MKN45 cells and to explore the mechanism of radiosensitization of erlotinib. Methods The growth inhibition effect of erlotinib and 5-fluorouracil on MKN45 cells was detected by MTT assay. The apoptosis of MKN45 cells treated with erlotinib, 5-Fu and combined radiation was detected by flow cytometry The effects of erlotinib, 5-Fu and combined radiation on Bax and Bcl-2 protein expression in MKN45 cells were detected by Western Blots. Results Both erlotinib and 5-Fu could inhibit the growth of MKN45 cells, which was positively correlated with drug concentration and time (P <0.01). Erlotinib group increased G2 / M, G0 / G1 phase cell ratio was the most obvious, compared with 5-Fu group was statistically significant (70.32 ± 0.65vs58.25 ± 0.42, P <0.01); apoptosis (19.21 ± 1.23 vs 13.11 ± 0.76, P <0.01). The expression of Bcl-2 protein was significantly decreased in the combined group of Erlotinib, while the expression of Bax The protein expression was significantly increased compared with other groups (P <0.01). Conclusions The mechanism of radiosensitization of erlotinib is related to the increase of the apoptosis rate of MKN45 cells, the ratio of G2 / M and G0 / G1 phase cells, the decrease of Bcl-2, the increase of Bax protein expression and the decrease of Bcl-2 / Bax ratio related.