论文部分内容阅读
目的 :观察核仁磷酸蛋白(nucleophosmin,NPM)和c-Src蛋白在耐药肺腺癌细胞中的表达及其与化疗敏感性之间的关系。方法 :通过顺铂浓度梯增渐进式给药法筛选出不同程度的肺腺癌耐药细胞,并将其接种于小鼠皮下。实验分为4组:对照组(小鼠皮下接种耐0.0μg/m L顺铂的Lewis肺癌细胞)、非耐药组(小鼠皮下接种耐0.0μg/m L顺铂的Lewis肺癌细胞)、低耐药组(小鼠皮下接种耐10.0μg/m L顺铂的Lewis肺癌细胞)和高耐药组(小鼠皮下接种耐20.0μg/m L顺铂的Lewis肺癌细胞)。对照组小鼠腹腔注射0.9%氯化钠溶液,非耐药组、低耐药组和高耐药组小鼠腹腔注射顺铂(4 mg/kg)进行化疗,观察小鼠皮下移植瘤的生长情况和肺转移情况,应用免疫组织化学法、蛋白质印迹法和实时荧光定量PCR法检测小鼠皮下移植瘤组织中NPM和c-Src蛋白及m RNA的表达。结果 :非耐药组、低耐药组和高耐药组小鼠皮下移植瘤的体积和质量均小于对照组(P值均<0.01),非耐药组、低耐药组和高耐药组的抑瘤率分别为73.23%、51.45%和32.74%。低耐药组和高耐药组小鼠皮下移植瘤组织中NPM和c-Src m RNA及蛋白的表达水平均高于非耐药组和对照组(P值均<0.01),高耐药组小鼠皮下移植瘤组织中NPM和c-Src m RNA及蛋白的表达水平均高于低耐药组(P值均<0.01),而非耐药组与对照组小鼠皮下移植瘤组织中NPM和c-Src mRNA及蛋白表达水平之间的差异无统计学意义(P值均>0.05)。结论 :NPM和c-Src在耐药的Lewis肺癌细胞中表达上调,可能与肺腺癌细胞对顺铂耐药有关。
OBJECTIVE: To observe the expression of nucleophosmin (NPM) and c-Src protein in drug-resistant lung adenocarcinoma cells and its relationship with chemosensitivity. Methods: Different doses of drug-resistant lung adenocarcinoma cells were screened by progressive administration of cisplatin concentration ladder and inoculated subcutaneously in mice. The experiment was divided into 4 groups: control group (mice were subcutaneously inoculated with Lewis lung cancer cells resistant to 0.0 μg / m L cisplatin), non-drug resistant mice (mice were subcutaneously inoculated with 0.06 μg / m L cisplatin Lewis lung cancer cells) Low-resistant group (Lewis lung cancer cells were subcutaneously inoculated with 10.0μg / m L cisplatin) and high-resistant group (mice were inoculated subcutaneously with Lewis lung cancer cells resistant to 20.0μg / m L cisplatin). Control mice were injected intraperitoneally with 0.9% sodium chloride solution, non-drug-resistant group, low-drug-resistant group and high-drug-resistant mice. The mice were injected cisplatin (4 mg / kg) The expression of NPM and c-Src protein and m RNA in subcutaneously transplanted tumor tissues were detected by immunohistochemistry, Western blotting and real-time fluorescence quantitative PCR. Results: The volume and quality of subcutaneously transplanted tumors in non-resistant group, low-resistant group and high-resistant group were significantly lower than those in control group (all P <0.01), non-resistant group, low-resistant group and high resistant The tumor inhibition rates were 73.23%, 51.45% and 32.74% respectively. The expression levels of NPM and c-Src m RNA and protein in subcutaneously transplanted tumor tissues of low-drug-resistant group and high-drug-resistant group were significantly higher than those of non-drug group and control group (all P <0.01) The expression levels of NPM and c-Src m RNA and protein in subcutaneously transplanted tumor tissues of mice were significantly higher than those in the low drug resistance group (all P <0.01), but the non-drug resistant group and the control group There was no significant difference in the expression of c-Src mRNA and protein (P> 0.05). CONCLUSION: NPM and c-Src are up-regulated in drug-resistant Lewis lung cancer cells, which may be related to the resistance of lung adenocarcinoma cells to cisplatin.