金复康口服液联合吉非替尼对H1975肺癌裸鼠移植瘤生长及有氧糖酵解的影响

Effect of Jinfukang Oral Liquid Combined with Gefitinib on the Growth and Aerobic Glycolysis of H1975 Lung Cancer Xenografts in Nude Mice

目的 观察金复康口服液联合吉非替尼对肺癌H1975细胞裸鼠移植瘤生长及有氧糖酵解的影响。方法 采用生物信息学分析HK2、PFKP、PKM2、LDHA在肺腺癌中的表达及对诊断和预后的影响;建立人肺腺癌H1975细胞荷瘤裸鼠模型后随机分为模型组、吉非替尼组(38 mg/kg)、金复康口服液组(25 g/kg生药)和联合用药组(38 mg/kg吉非替尼+25 g/kg金复康口服液生药),每组6只。每周测量移植瘤体积及裸鼠体质量2次,给药21日后取肿瘤组织称重并计算相对肿瘤增殖率及抑瘤率;采用RT-PCR、Western Blot法检测各组移植瘤组织HK2、PFKP、PKM2、LDHA的m RNA及蛋白表达;微量法检测糖酵解酶活性、ATP及乳酸含量。结果 生物信息学分析显示PFKP、PKM2和LDHA在肺腺癌中高表达,对肺腺癌的诊断和预后判断具有一定参考意义。与模型组及单药组比较,联合用药组肿瘤体积及肿瘤质量降低(P<0.01),抑瘤率提高(P<0.01)。与模型组比较,联合用药组HK2、PFKP及PKM2的m RNA及蛋白表达下调(P<0.01),LDHA蛋白表达无明显变化,移植瘤内HK、PFK活性下调,ATP和乳酸含量降低(P<0.01)。结论 复康口服液能增强耐药H1975细胞荷瘤裸鼠对吉非替尼的敏感性延缓肿瘤生长,其机制可能是通过下调HK2、PFKP和PKM2水平,减少ATP及乳酸生成,抑制有氧糖酵解实现的。

Objective To observe the effect of Jinfukang Oral Liquid combined with gefitinib on the growth and aerobic glycolysis in nude mice xenografts model of H1975 lung cancer. Methods Bioinformatics was used to analyze the expressions of HK2,PFKP,PKM2,and LDHA in lung adenocarcinoma and their effects on diagnosis and prognosis. After the establishment of human lung adenocarcinoma H1975 cells tumor-bearing nude mice model, they were randomly divided into model group,gefitinib group(38 mg/kg),Jinfukang Oral Liquid group(25 g/kg crude drugs) and combination group(38 mg/kg gefitinib and 25 g/kg Jinfukang Oral Liquid crude drugs),with 6 nude mice in each group. Tumor volume and body mass of nude mice were measured twice a week, and tumor tissues were weighed and the relative tumor proliferation rate and tumor suppression rate were calculated after 21 days of drug administration.RT-PCR and Western Blot were used to detect the mRNA and protein expression of HK2,PFKP,PKM2 and LDHA in the transplanted tumor tissues.The glycolytic enzyme activity,ATP and lactic acid contents were detected by micromethod. Results Bioinformatics analysis showed that PFKP,PKM2 and LDHA were highly expressed in lung adenocarcinoma,which had certain reference significance for the diagnosis and prognosis of lung adenocarcinoma.Compared with the model group and single drug group,the tumor volume and mass of the combined group were decreased(P<0.01),and the tumor inhibition rate was increased(P<0.01).The mRNA and protein expression of HK2,PFKP and PKM2 in the combination group were significantly down-regulated compared with those in the model group(P<0.01),while the protein expression of LDHA was not significantly changed.The activities of HK,PFK and the contents of ATP and lactic acid in the combined group were significantly lower than those in the model group(P<0.01). Conclusion Jinfukang Oral Liquid can enhance the sensitivity of drug-resistant H1975 cell bearing nude mice to gefitinib and inhibit tumor growth,which may be achieved by down-regulating the expression of HK2,PFKP and PKM2,reducing ATP and lactic acid production,and inhibiting aerobic glycolysis.