健脾温肾方对皮下移植瘤术后模型裸鼠肺转移瘤体中miRNAs及相关靶基因mRNA表达的影响

Effects of Jianpi Wenshen Fang(健脾温肾方) on Expressions of miRNAs and Related Target Genes mRNA in Lung Metastases of Nude Mice after Subcutaneous Transplantation

目的探讨健脾温肾方对肺癌术后复发转移的预防作用及可能机制。方法建立NCI-H460-Luci细胞皮下移植瘤术后裸鼠模型,随机分为健脾温肾组、化疗组、对照组,每组16只。健脾温肾组术后第1天开始给予健脾温肾方溶液(1.1 g/ml)0.3 ml/d灌胃,每日1次,干预13周;化疗组给予紫杉醇注射液15 mg/kg,每隔5天腹腔注射1次,共用3次;对照组给予生理盐水0.3 ml/d灌胃,每日1次,干预13周。每周1次至术后13周,采用活体成像观察裸鼠肿瘤转移情况,分析各组术后无瘤转移生存时间;术后第6周,采用实时荧光定量PCR法检测瘤体中miR-18a-5p、miR182-5p、miR21、miR106b-5p及相关靶基因乳腺癌易感基因1(BRCA1)、重组人磷酸酶及张力蛋白同源体(PTEN)、P53肿瘤蛋白(TP53)、表皮生长因子受体(EGFR)mRNA表达量。结果与对照组比较,健脾温肾组、化疗组裸鼠瘤体中miR-18a-5p、miR182-5p表达降低,BRCA1 mRNA表达升高(P<0.05或P<0.01);化疗组裸鼠瘤体中miR21、miR106b-5p表达亦较对照组降低(P<0.05或P<0.01)。各组裸鼠瘤体中EGFR、PTEN、TP53 mRNA表达差异均无统计学意义(P>0.05)。健脾温肾组和对照组无瘤转移生存时间比较差异具有统计学意义(P<0.05),化疗组与健脾温肾组比较差异无统计学意义(P>0.05)。结论健脾温肾方在预防肺癌术后转移方面有一定作用,其机制可能与调节miRNAs及相关靶基因mRNA有关。

Objective To investigate the preventive effect of Jianpi Wenshen Fang （健脾温肾方） on the recur- rence and metastasis of lung cancer and its possible mechanism. Methods NCI-H460-Luci cells were used to establish a nude mouse model after subcutaneous transplantation and were randomly divided into invigorating spleen and warming kidney group, chemotherapy group and control group, with 16 mice in each group. On the first postoperative day, the invigorating spleen and warming kidney group wag given intragastric administration of Jianpi Wenshen Fang （1.1 g/ml） 0. 3 ml/d, once a day for 13 weeks; the chemotherapy group was given paclitaxel 15 mg/kg. The rats were intraperitoneally injected once every 5 days totally for 3 times. The control group was given normal saline 0. 3 ml/d orally once a day for 13 weeks. Until 13 weeks after operation, tumor metastasis of nude mice was observed by in vivo imaging once a week. The survival time of tumor-free metastasis in each group was analyzed. At the 6th week after operation, miR-18a-5p, miR182-Sp, miR21, miR106b-Sp and related target genes breast cancer suscepti- bility gene 1 （ BRCA1 ）, recombinant human phosphatase and tensin homology （ PTEN）, P53 tumor protein （ TP53 ）, epidermal growth factor receptor （EGFR） mRNA expression levels were detected by real-time fluorescence quantita- tive PCR. Results Compared with the control group, the expression of miR-18a-5p and miR182-5p decreased and the expression of BRCA1 mRNA increased in the nude mice of the invigorating spleen and warming kidney group and the chemotherapy group （P 〈0.05 or P 〈0. 01 ） ; The expression of miR21 and miR106b-Sp in tumors of nude mice in the chemotherapy group was also lower than that in control group （P 〈 0. 05 or P 〈 0. 01 ）, There was no significant difference in the expression of EGFR, PTEN and TP53 mRNA in the tumors of each group （ P 〉 0.05 ）. The diffe- rence in the survival time of tumor-free metastasis between the invigorating spleen and warming kidney group and the control group was statistically significant （P 〈 0. 05 ）, and there was no significant difference between the chemothera- py group and the invigorating spleen and warming kidney group （ P 〉 0. 05 ）. Conclusion Jianpi Wenshen Fang has a certain role in preventing postoperative metastasis of lung cancer, and its mechanism may be related to regulating miRNAs and related target gene mRNA.