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目的探讨环巴胺联合卡铂对人肺腺癌A549细胞的抑制作用及其可能机制。方法采用MTT法检测环巴胺和卡铂单独或联合给药对人肺腺癌A549细胞增殖的抑制作用;建立人肺腺癌A549裸鼠异种移植模型,分为对照组、卡铂组(CBP组,10 mg/kg)、环巴胺高剂量组1(CYCPH 1组,2.5 mg/kg)、环巴胺高剂量组2(CYCPH 2组,2.5 mg/kg,分组后第6天给药)、环巴胺中剂量组(CYCPM组,1.0 mg/kg)、环巴胺低剂量组(CYCPL组,0.5 mg/kg)和联合给药组(卡铂同CBP组+环巴胺同CYCPH 2组),每周2次测量肿瘤体积,并根据相对肿瘤体积计算肿瘤抑制率,称重裸鼠去瘤后体重;采用免疫组化检测细胞增殖和凋亡标志物Ki-67和TUNEL的表达;蛋白质印迹法(Western blot)检测肿瘤组织中Bcl-2以及Bax的表达情况。结果体外抗肿瘤作用结果显示,环巴胺和卡铂单独给药组细胞增殖抑制率分别为49.87%和56.38%,联合给药组的增殖抑制率为79.11%。体内抗肿瘤作用结果显示,与对照组比较,CBP组、CYCPH 2组和联合给药组的裸鼠去瘤后体重明显降低,差异有统计学意义(P<0.05,P<0.01);CYCPL组、CYCPM组、CYCPH 1组和CYCPH 2组的肿瘤抑制率分别为18.26%、34.45%、47.60%和40.04%,联合给药组的肿瘤抑制率为64.30%,明显高于各单独给药组(P<0.01)。免疫组化结果显示,与卡铂和环巴胺单独给药组相比,联合给药组TUNEL的表达更高,Ki-67的表达更低。Western blot结果显示,与卡铂和环巴胺单独给药组相比,联合用药组Bcl-2表达明显降低,Bax表达明显提高。结论环巴胺与卡铂联用能够有效地抑制人肺腺癌A549细胞生长,作用机制可能与调节凋亡相关蛋白的表达有关。
Objective To investigate the inhibitory effect of cyclopamine combined with carboplatin on human lung adenocarcinoma A549 cells and its possible mechanism. Methods The inhibitory effect of cyclopamine and carboplatin alone or in combination on the proliferation of human lung adenocarcinoma A549 cells was detected by MTT assay. The human lung adenocarcinoma A549 xenograft model was established and divided into control group, carboplatin group (CBP (CYCPH 1, 2.5 mg / kg), cyclopamine high dose group 2 (CYCPH 2, 2.5 mg / kg, dosing 6 days after grouping) (CYCPM group, 1.0 mg / kg), low dose cyclopamine group (CYCPL group, 0.5 mg / kg) and combination group (carboplatin and CBP group + cyclopamine with CYCPH 2 groups). The tumor volume was measured twice a week, and the tumor inhibition rate was calculated according to the relative tumor volume. The tumor weight of nude mice was weighed. The expression of Ki-67 and TUNEL, a marker of cell proliferation and apoptosis, was detected by immunohistochemistry Western blot was used to detect the expression of Bcl-2 and Bax in tumor tissue. Results The anti-tumor effect in vitro showed that the cell proliferation inhibition rates of cyclopamine and carboplatin alone group were 49.87% and 56.38%, respectively. The proliferation inhibition rate of the combination group was 79.11%. The antitumor effect in vivo showed that the body weight of nude mice in CBP group, CYCPH 2 group and combination group was significantly lower than that in control group (P <0.05, P <0.01); CYCPL group The tumor inhibition rates of CYCPM group, CYCPH 1 group and CYCPH 2 group were 18.26%, 34.45%, 47.60% and 40.04%, respectively. The tumor inhibition rate of combined group was 64.30%, which was significantly higher than that of the other groups P <0.01). The results of immunohistochemistry showed that TUNEL expression was higher and the expression of Ki-67 was lower in combination group than carboplatin and cyclopamine alone. Western blot results showed that compared with carboplatin and cyclopamine alone, the combination group Bcl-2 expression was significantly lower, Bax expression was significantly increased. Conclusions The combination of cyclopamine and carboplatin can effectively inhibit the growth of human lung adenocarcinoma A549 cells. The mechanism may be related to the regulation of the expression of apoptosis-related proteins.