HSP27在食管鳞状细胞癌KYSE150细胞中的功能
|
摘要
目的研究热休克蛋白27(HSP27)对食管鳞状细胞癌(ESCC)细胞迁移、侵袭、增殖的作用。方法用Western blot技术检测5种食管癌细胞系(KYSE30、KYSE150、KYSE510、TE-1、Eca9706)中HSP27的本底表达情况。用慢病毒转染HSP27的短发夹RNA(short hairpin RNA,shRNA)至KYSE150细胞系,实验共分5组:Empty vector组(未转染的KYSE150细胞),Negative control组(转染随机序列),3组实验组(分别转染shRNA-HSP27-1、shRNA-HSP27-2、shRNA-HSP27-3)。用噻唑蓝比色法(MTT)、Transwell侵袭实验以及细胞划痕实验检测HSP27不同表达水平对食管鳞状细胞癌细胞增殖、侵袭及迁移能力的影响。结果 5种食管癌细胞系中HSP27本底表达量由高到低分别是:KYSE150、KYSE510、KYSE30、Eca9706、TE-1。与Empty vector组和Negative control组相比,HSP27敲低的实验组中食管鳞状细胞癌细胞的增殖、侵袭、迁移能力明显受抑制(P<0.05),差异有统计学意义。结论 HSP27在食管鳞状细胞癌中对肿瘤细胞的增殖、侵袭、迁移能力有促进作用,研究结果为食管癌的临床分子靶向治疗提供了新的研究方向。
Objective To investigate the effects of heat shock protein 27(HSP27) on migration, invasion, proliferation and apoptosis of esophageal squamous cell carcinoma(ESCC) cells. Methods The background expression of HSP27 in five esophageal cancer cell lines(KYSE30, KYSE150, KYSE510, TE-1, Eca9706) was detected by Western blot. HSP27 short hairpin RNA(shRNA) was transfected into the KYSE150 cell line with lentivirus. The experiment was divided into five groups: empty vector group(untransfected), negative control group(transfected random sequence), and three experimental groups(transfected shRNA, respectively sh-HSP27-1, shRNA-HSP27-2, shRNA-HSP27-3). The effects of different expression of HSP27 on proliferation, invasion, migration and apoptosis of esophageal squamous carcinoma cells were detected by MTT, transwell assay, cell scratch assay and flow cytometry(FCM). Results The high-to-low expression levels of HSP27 in esophageal cancer cell lines were: KYSE150, KYSE510, KYSE30, Eca9706, TE-1. Compared with the Empty vector group and the Negative control group, the proliferation, invasion and migration of esophageal cancer cells in the three groups with HSP27 knockdown were significantly inhibited significantly(P<0.05), and the apoptosis ability was enhanced significantly(P<0.05). Conclusion HSP27 promotes the proliferation, invasion and migration of tumor cells in esophageal squamous cell carcinoma and inhibits apoptosis. The results provide a potential target for clinical molecular targeted therapy of esophageal cancer.
Objective To investigate the effects of heat shock protein 27(HSP27) on migration, invasion, proliferation and apoptosis of esophageal squamous cell carcinoma(ESCC) cells. Methods The background expression of HSP27 in five esophageal cancer cell lines(KYSE30, KYSE150, KYSE510, TE-1, Eca9706) was detected by Western blot. HSP27 short hairpin RNA(shRNA) was transfected into the KYSE150 cell line with lentivirus. The experiment was divided into five groups: empty vector group(untransfected), negative control group(transfected random sequence), and three experimental groups(transfected shRNA, respectively sh-HSP27-1, shRNA-HSP27-2, shRNA-HSP27-3). The effects of different expression of HSP27 on proliferation, invasion, migration and apoptosis of esophageal squamous carcinoma cells were detected by MTT, transwell assay, cell scratch assay and flow cytometry(FCM). Results The high-to-low expression levels of HSP27 in esophageal cancer cell lines were: KYSE150, KYSE510, KYSE30, Eca9706, TE-1. Compared with the Empty vector group and the Negative control group, the proliferation, invasion and migration of esophageal cancer cells in the three groups with HSP27 knockdown were significantly inhibited significantly(P<0.05), and the apoptosis ability was enhanced significantly(P<0.05). Conclusion HSP27 promotes the proliferation, invasion and migration of tumor cells in esophageal squamous cell carcinoma and inhibits apoptosis. The results provide a potential target for clinical molecular targeted therapy of esophageal cancer.
引文
[1] FERLAY J, SOERJOMATARAM I, DIKSHIT R, et al. Cancer incidence and mortality worldwide:sources,methods and major patterns in GLOBOCAN 2012 [J].Int J Cancer,2015, 136(5):359-386.
[2] 鲁智豪.ASCO-GI 2019速递—食管癌篇[J].肿瘤综合治疗电子杂志,2019(1):117-120.
[3] MUNARI F F, CRUVINEL C A, LACERDA C F, et al. PIK3CA mutations are frequent in esophageal squamous cell carcinoma associated with chagasic megaesophagus and are associated with a worse patient outcome[J].Infect Agent Cancer, 2018,13:43.
[4] QI Y J, CHAO W X, CHIU J F. An overview of esophageal squamous cell carcinoma proteomics[J].Proteomics,2012,11:3129-3137.
[5] MARIETTE C, MARKAR S R, DABAKUYO T S, et al. Hybrid minimally invasive esophagectomy for esophageal cancer[J].N Engl J Med,2019,380(2):152-162.
[6] CALDERWOOD S K, GONG J. Heat shock proteins promote cancer:It's a protection racket[J].Trends Biochem Sci,2016,41(4):311-323.
[7] 蔡洪庆.胶质瘤中HSP27、p-HSP27和MCM6表达与临床意义[D].北京:北京协和医学院,2018.
[8] KAYSER J, HASLBECK M, DEMPFLE L, et al. The small heat shock protein Hsp27 affects assembly dynamics and structure of keratin intermediate filament networks[J].Biophys J, 2013,105:1778-1785.
[9] WANG W, MA J, LU J, et al. Circ0043898 acts as a tumor inhibitor and performs regulatory effect on the inhibition of esophageal carcinoma[J]. Cancer Biol The,2018,19(12): 1117-1127.
[10] CHATTERJEE S, BURNS T F. Targeting heat shock proteins in cancer:a promising therapeutic approach[J].Int J Mol Sci,2017,18(9):1978.
[11] SAINI J, SHARMA P K. Clinical, prognostic and therapeutic significance of heat shock proteins in cancer[J].Curr Drug Targets,2018,19(13):1478-1490.
[12] TRINKLEIN N D, CHEN W C, KINGSTON R E, et al. Transcriptional regulation and binding of heat shock factor 1 and heat shock factor 2 to 32 human heat shock genes during thermal stress and differentiation[J].Cell Stress Chaperon,2004,9:21-28.
[13] YE H, HUANG H, CAO F, et al. HSPB1 enhances SIRT2-Mediated G6PD activation and promotes glioma cell proliferation[J].PLoS One,2016,11(10):e0164285.
[14] HUANG C S, HUANG C Y, LEE C H, et al. IGFBP2 plays an important role in heat shock protein 27-mediated cancer progression and metastasis[J].Oncotarget,2017,8(33):54978-54992.
[15] GE H, DU J, XU J, et al. SUMOylation of HSP27 by small ubiquitin-like modifier 2/3 promotes proliferation and invasion of hepatocellular carcinoma cells[J].Cancer Biol Ther,2017,18(8):552-559.
[2] 鲁智豪.ASCO-GI 2019速递—食管癌篇[J].肿瘤综合治疗电子杂志,2019(1):117-120.
[3] MUNARI F F, CRUVINEL C A, LACERDA C F, et al. PIK3CA mutations are frequent in esophageal squamous cell carcinoma associated with chagasic megaesophagus and are associated with a worse patient outcome[J].Infect Agent Cancer, 2018,13:43.
[4] QI Y J, CHAO W X, CHIU J F. An overview of esophageal squamous cell carcinoma proteomics[J].Proteomics,2012,11:3129-3137.
[5] MARIETTE C, MARKAR S R, DABAKUYO T S, et al. Hybrid minimally invasive esophagectomy for esophageal cancer[J].N Engl J Med,2019,380(2):152-162.
[6] CALDERWOOD S K, GONG J. Heat shock proteins promote cancer:It's a protection racket[J].Trends Biochem Sci,2016,41(4):311-323.
[7] 蔡洪庆.胶质瘤中HSP27、p-HSP27和MCM6表达与临床意义[D].北京:北京协和医学院,2018.
[8] KAYSER J, HASLBECK M, DEMPFLE L, et al. The small heat shock protein Hsp27 affects assembly dynamics and structure of keratin intermediate filament networks[J].Biophys J, 2013,105:1778-1785.
[9] WANG W, MA J, LU J, et al. Circ0043898 acts as a tumor inhibitor and performs regulatory effect on the inhibition of esophageal carcinoma[J]. Cancer Biol The,2018,19(12): 1117-1127.
[10] CHATTERJEE S, BURNS T F. Targeting heat shock proteins in cancer:a promising therapeutic approach[J].Int J Mol Sci,2017,18(9):1978.
[11] SAINI J, SHARMA P K. Clinical, prognostic and therapeutic significance of heat shock proteins in cancer[J].Curr Drug Targets,2018,19(13):1478-1490.
[12] TRINKLEIN N D, CHEN W C, KINGSTON R E, et al. Transcriptional regulation and binding of heat shock factor 1 and heat shock factor 2 to 32 human heat shock genes during thermal stress and differentiation[J].Cell Stress Chaperon,2004,9:21-28.
[13] YE H, HUANG H, CAO F, et al. HSPB1 enhances SIRT2-Mediated G6PD activation and promotes glioma cell proliferation[J].PLoS One,2016,11(10):e0164285.
[14] HUANG C S, HUANG C Y, LEE C H, et al. IGFBP2 plays an important role in heat shock protein 27-mediated cancer progression and metastasis[J].Oncotarget,2017,8(33):54978-54992.
[15] GE H, DU J, XU J, et al. SUMOylation of HSP27 by small ubiquitin-like modifier 2/3 promotes proliferation and invasion of hepatocellular carcinoma cells[J].Cancer Biol Ther,2017,18(8):552-559.