益气除痰方对肺癌移植鼠内质网分子伴侣蛋白GRP94表达的影响.docVIP

益气除痰方对肺癌移植鼠内质网分子伴侣蛋白GRP94表达的影响 　　摘要：目的 观察益气除痰方对脾虚Lewis肺癌移植鼠内质网分子伴侣蛋白GRP94表达的影响，探讨其抗癌的可能作用机制。方法 采用苦寒泻下加饥饱失常方式建立脾虚小鼠模型，再将无血清培养基制备的癌细胞悬液接种至脾虚C57BL/6小鼠背侧，制作脾虚Lewis肺癌小鼠模型。将48只成模小鼠随机分为模型组、化疗组、联合用药组及益气除痰方低、中、高剂量组，每组8只。各给药组给予相应药物干预。给药14 d后处死小鼠，检测肿瘤体积和质量；HE染色观察各组移植瘤组织形态，免疫组化、Western blot和RT-PCR检测肿瘤组织GRP94蛋白和基因表达。结果 与模型组比较，化疗组、联合用药组及益气除痰方中、高剂量组肿瘤体积、肿瘤质量明显降低（P0.05，P0.01），肿瘤细胞大量坏死，GRP94蛋白表达明显降低（P0.01）；与化疗组比较，联合用药组GRP94蛋白表达明显降低（P0.01）。结论 益气除痰方联合化疗可有效治疗Lewis肺癌小鼠，其机制可能与其下调内质网分子伴侣蛋白GRP94表达、逆转肿瘤细胞自身保护机制相关。 　　关键词：益气除痰方；肺癌；脾虚证；GRP94；小鼠 　　DOI：10.3969/j.issn.1005-5304.2016.12.015 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）12-0059-05 　　Effects of Yiqi Chutan Formula on GRP94 of Endoplasmic Reticulum Molecular Chaperone in Mice with Lung Cancer WEI Yu-lin， WANG Shu-mei， LIU Qi-ou （Chongqing Medical University， Chongqing 400016， China） 　　Abstract：Objective To investigate the effects of Yiqi Chutan Formula on expression of GRP94 of endoplasmic reticulum molecular chaperone in mice with lung cancer. Methods The binding methods of diarrhea of bitter and cold and abnormal of starvaion were used to establish spleen-deficiency mice models. Cancer cell suspension prepared by serum-free medium was inoculated to the back of C57BL/6 mice to establish spleen-deficiency Lewis lung cancer models. 48 successfully modeling spleen-deficiency Lewis lung cancer mice were randomly divided into model group， chemotherapy group， Yiqi Chutan Formula low-， medium-， and high-dose groups， and combined medication group， with 8 mice in each group. Each medication group was intervened with relevant medicine. The mice were killed after 14 days of medication. The size and weight of tumor in mice were measured； morphological changes of transplanted tumor were observed by HE staining method； the protein and gene expressions of GRP94 in tumor tissues were tested by immunohistochemistry， Western blot and real-time fluorescence quantitative PCR. Results Compared with model group， the tumor volume and weight of chem