葡萄糖调节蛋白78对非小细胞肺癌患者吉西他滨化疗敏感性的影响

doi:10.13481/j.1671-587x.20190321

摘要/Abstract

摘要： 目的：探讨葡萄糖调节蛋白78（GRP78）表达与非小细胞肺癌（NSCLC）患者吉西他滨化疗的关系及GRP78对肺腺癌SPCA1细胞活力和吉西他滨化疗敏感性的影响，并阐述其作用机制。方法：选取32例NSCLC患者的癌组织石蜡标本，采用免疫组织化学染色检测NSCLC癌组织中GRP78阳性表达率。选取GRP78高表达的SPCA1细胞作为研究对象。采用RNA干扰技术在SPCA1细胞中下调GRP78的表达水平（干扰组），并设对照组（给予shNC）；MTT法检测各组SPCA1细胞活力。实验分为阴性对照组、干扰组、对照＋吉西他滨组和干扰＋吉西他滨组，克隆形成实验检测各组SPCA1细胞克隆形成率，Western blotting法检测各组SPCA1细胞中Akt、p-Akt、PI3K和p-PI3K的表达量。结果：免疫组织化学染色法检测，与经吉西他滨治疗缓解的NSCLC患者癌组织比较，经吉西他滨治疗未缓解NSCLC患者癌组织中GRP78阳性表达率明显升高（P<0.05）。MTT法检测，与对照组比较，干扰组细胞活力明显降低（P<0.05）。克隆形成实验，与阴性对照组比较，干扰组SPCA1细胞克隆形成率明显降低（P<0.05），干扰＋吉西他滨组SPCA1细胞克隆形成率降低更加明显（P<0.05）。Western blotting法检测，与阴性对照组比较，干扰组细胞中p-Akt、p-PI3K表达量明显降低，干扰＋吉西他滨组细胞中p-Akt和p-PI3K表达量降低更加明显。结论：干扰GRP78可以增加吉西他滨化疗的敏感性，GRP78可能通过PI3K/Akt途径降低NSCLC患者对吉西他滨的敏感性。

关键词: 癌,非小细胞肺, 葡萄糖调节蛋白78, 吉西他滨, 化疗敏感性

Abstract: Objective:To investigate the relationship between the expression of glucose-regulated protein 78 (GRP78) and gemcitabine chemotherapy in the patients with non-small cell lung cancer (NSCLC) and the effects of GRP78 on the viability of lung adenocarcinoma SPCA1 cells and the chemosensitivity of gemcitabine, and to elucidate its mechanisms. Methods:The positive expression rates of GRP78 in 32 cases of cancer tissue of the NSCLC patients were detected by immunohistochemical staining. The SPCA1 cells with high expression of GRP78 were selected as the subjects. RNA interference technique was used to down-regulate the expression of GRP78 in SPCA1 cells(interference group) and the cells treated with shNC were used as control group. MTT assay was used to detect the viabilities of SPCA1 cells in various groups.Negative control group,interference group,control+gemcitabine group,and interference+gemcitabine group were set up; colone formation assay was used to detect the colone formation rates of SPCA1 cells in various groups; Western blotting method was used to detect the expression amount of Akt, p-Akt, PI3K, and p-PI3K in the SPCA1 cells in various groups. Results:The immunohistochemical staining results showed that the positive expression rate of GRP78 in cancer tissue in the remission NSCLC patients was significantly higher than that in the no-remission NSCLC patients after treated with gemcitabine (P<0.05). The MTT assay results showed that compared with negative control group, the viability of SPCA1 cells in interference group was decreased significantly (P<0.05),and the viability of SPCA1 cells in interference+gemcitabine group was significantly decresed(P<0.05). The Western blotting results showed that compared with negative control group, the expression amounts of p-Akt and p-PI3K in the SPCA1 cells in interference group were decreased, and the expression amounts of p-Akt and p-PI3K in the SPCA1 cells in interference+gemcitabine group were decreased significantly. Conclusion:Interference of GRP78 may increase the sensitivity of gemcitabine to chemotherapy,and GRP78 may reduce the sensitivity of NSCLC patients to gemcitabine through PI3K/Akt pathway.

Key words: cancer,non-small cell lung, glucose-regulated protein 78, gemcitabine, chemosensitivity

中图分类号:

R734.2

王志静, 苏荣健, 杜晓媛. 葡萄糖调节蛋白78对非小细胞肺癌患者吉西他滨化疗敏感性的影响[J]. 吉林大学学报(医学版), 2019, 45(03): 595-600.

WANG Zhijing, SU Rongjian, DU Xiaoyuan. Effects of GRP78 on sensibility of gemcitabine on patients with NSCLC[J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 595-600.

参考文献

[1] ULIVI P, ZOLI W, CAPELLI L, et al. Target therapy in NSCLC patients:Relevant clinical agents and tumour molecular characterisation[J].Mol Clin Oncol, 2013, 1(4):575-581.
[2] FAES S, DORMOND O. PI3K and AKT:unfaithful partners in cancer[J]. Int J Mol Sci, 2015, 16(9):21138-21152.
[3] TUYA N, WANG Y, TONG L, et al. Trichosanthin enhances the antitumor effect of gemcitabine in non-small cell lung cancer via inhibition of the PI3K/AKT pathway[J]. Exp Ther Med, 2017, 14(6):5767-5772.
[4] FU R, YANG P, WU H L, et al. GRP78 secreted by colon cancer cells facilitates cell proliferation via PI3K/Akt signaling[J]. Asian Pac J Cancer Prev, 2014, 15(17):7245-7249.
[5] GIFFORD J, HILL R. GRP78 influences chemoresistance and prognosis in cancer[J]. Current Drug Targets, 2018, 19(6):701-708.
[6] 王立文, 沈晓洁, 林彬. 非小细胞肺癌中lncRNA MVIH与吉西他滨耐药的相关性研究[J]. 重庆医学, 2017, 46(7):946-948.
[7] CHEN Y, HUANG Y, CHEN D M, et al. RRM1 expression and the clinicopathological characteristics of patients with non-small cell lung cancer treated with gemcitabine[J]. OncoTargets Ther, 2018, 11:5579-5589.
[8] XU C, YU Y, DING F. Microarray analysis of circular RNA expression profiles associated with gemcitabine resistance in pancreatic cancer cells[J]. Oncol Rep, 2018, 40(1):395-404.
[9] XIONG G, HUANG H, FENG M, et al. MiR-10a-5p targets TFAP2C to promote gemcitabine resistance in pancreatic ductal adenocarcinoma[J]. J Exp Clin Cancer Res, 2018, 37(1):76-87.
[10] WANG Y, SUI Y, YAN K, et al. BRD4 promotes pancreatic ductal adenocarcinoma cell proliferation and enhances gemcitabine resistance[J]. Oncol Rep, 2015, 33(4):1699-1706.
[11] LEE J, YOON Y, LEE S. GRP78 regulates apoptosis, cell survival and proliferation in 5-Fluorouracil-resistant SNUC5 colon cancer cells[J]. Anticancer Res, 2017, 37(9):4943-4951.
[12] LIZARDO M, MORROW J, MILLER T, et al. Upregulation of glucose-regulated protein 78 in metastatic cancer cells is necessary for lung metastasis progression[J]. Neoplasia, 2016, 18(11):699-710.
[13] WANG Y, WANG J H, ZHANG X L, et al. Endoplasmic reticulum chaperone glucose-regulated protein 78 in gastric cancer:An emerging biomarker[J]. Oncol Lett, 2018, 15(5):6087-6093.
[14] DANESHMAND S, QUEK M L, LIN E, et al. Glucose regulated protein GRP78 is up-regulated in prostate cancer and correlates with recurrence and survival[J]. Hum Pathol, 2007, 38(10):1547-1552.
[15] RUI L, GU Y, HE W, et al. Secreted GRP78 activates EGFR-SRC-STAT3 signaling and confers the resistance to sorafeinib in HCC cells[J]. Oncotarget, 2017, 8(12):19354-19364.
[16] CHIOU J F, TAI C J, HUANG M T, et al. Glucose-regulated protein 78 is a novel contributor to acquisition of resistance to sorafenib in hepatocellular carcinoma[J]. Ann Surg Oncol, 2010, 17(2):603-612.
[17] JENIFER B, HUANG W, ZELENIAK A E. Expression of GRP78 master regulator of the unfolded protein response increases chemoresistance in pancreatic ductal adenocarcinoma[J]. Am Associat Cancer Res, 2016, 15(5):1043-1052.
[18] ZHANG Z, ZHANG M, LIU H, et al. AZD9291 promotes autophagy and inhibits PI3K/Akt pathway in NSCLC cancer cells[J]. J Cell Biochem, 2018, 120(1):756-767.
[19] CHEN Y, HUANG W, SUN W, et al. LncRNA MALAT1 promotes cancer metastasis in osteosarcoma via activation of the PI3K-Akt signaling pathway[J]. Cell Physiol Biochem, 2018, 51(3):1313-1326.
[20] 胡丽娜, 彭兴春, 郭显智, 等. 抑制Notch和PI3K/Akt信号通路对食管腺癌细胞增殖、侵袭及迁移的影响[J]. 肿瘤防治研究, 2016, 43(8):653-658.
[21] ZHENG Z, ZHOU X, CAI Y, et al. TEKT4 promotes papillary thyroid cancer cell proliferation, colony formation, and metastasis through activating PI3K/Akt pathway[J]. Endocr Pathol, 2018, 29(4):310-316.
[22] 苏宝安,许之晨,李艾恩.3.0T MR弥散加权成像评价中晚期食管鳞癌放化疗的早期疗效[J].中国医学物理学杂志,2018,35(12):1452-1456.