ERBB3表达下调对宫颈癌细胞增殖迁移和侵袭的影响
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摘要
目的探讨erb-b2受体酪氨酸激酶3(ERBB3)在调节癌细胞的迁移和侵袭及其在宫颈癌的发生和进展过程中的作用。方法收集2014年4月—2016年12月期间在本院接受治疗的25例宫颈鳞状细胞癌患者和25例宫颈腺癌患者。通过荧光定量PCR(qRT-PCR)检测了宫颈鳞状细胞癌和子宫颈腺癌患者体内肿瘤组织和癌旁组织中ERBB3 mRNA的表达水平。用siRNA转染宫颈鳞状细胞癌细胞沉默ERBB3的表达后,应用Transwell细胞迁移实验检测细胞迁移和侵袭能力,利用CCK-8检测细胞增殖,应用Western blotting检测MTK-1蛋白的表达变化。结果宫颈鳞状细胞癌和宫颈腺癌患者癌组织中ERBB3 mRNA水平均低于其正常组织(P=0.00,P=0.00)。HPV感染对ERBB3蛋白在宫颈鳞状细胞癌细胞系的表达水平为10.30±0.07(HPV阳性)、7.99±0.75(HPV阴性)和正常宫颈细胞系中的的表达水平为3.02±0.47(HPV阳性)、3.16±0.22(HPV阴性),差异均无统计学意义(P均>0.05)。ERBB3 si RNA沉默后宫颈鳞状细胞癌细胞SiHa和C33A的增殖(0.20±0.10,0.16±0.08)、迁移(64.87±0.26,55.02±0.06)和侵袭(49.99±0.06,62.80±0.19)能力显著低于正常宫颈细胞系中SiHa和C33A的增殖(0.86±0.49,0.60±0.34)、迁移(99.99±0.31,100.07±0.23)和侵袭(99.89±0.35,99.89±0.14)能力,差异具有统计学意义(P<0.05)。ERBB3通过si RNA沉默后MTK-1蛋白的表达水平也显着降低。结论 ERBB3的下调很可能通过抑制MTK-1的表达来抑制宫颈癌细胞的增殖、迁移和侵袭,从而抑制肿瘤的发展。
Objective To explore the role if erb-b2 receptor tyrosine kinase 3(ERBB3) in regulating the migration and invasion of cancer cells, and its role in the occurrence and progression of cervical cancer. Methods Totally 25 patients with cervical squamous cell carcinoma and 25 patients with cervical adenocarcinoma were enrolled who were treated in our center from April 2014 to December 2016. Expression level of ERBB3 in patients with cervical squamous cell carcinoma and cervical adenocarcinoma was detected by qRT-PCR. After ERBB3 siRNA silencing on cervical squamous cell, Transwell experiment was used to analyze the capability of migration and invasion, and CCK-8 kits was used to analyze the capability of proliferation, and Western blotting was used to detect S-methyl-5-thioribose kinase 1(MTK-1) protein level. Results qRT-PCR results showed that the expression levels of ERBB3 decreased in cervical squamous cell carcinoma and cervical adenocarcinoma were lower than those in the normal cervical cell lines(P=0.00, P=0.00). The expression level of HPV infection to ERBB3 protein in cervical squamous cell carcinoma cell lines was 10.30±0.07(HPV positive), 7.99 ±0.75(HPV negative) and normal cervical cell lines were 3.02±0.47(HPV positive) and 3.16±0.22(HPV negative), with no statistical significance(P>0.05). After ERBB3 si RNA silencing, the capabilities of SiHa and C33 A in cervical squamous cell carcinoma in proliferation(0.20±0.10, 0.16±0.08), migration(64.87±0.26, 55.02±0.06), and invasion(49.99±0.06, 62.80±0.19) were significantly lower than those in normal cervical cell lines(P<0.05). In addition, the expression level of MTK-1 protein was also significantly reduced after MTK-1 siRNA silencing. Conclusion Down-regulation of ERBB3 can decrease the proliferation, migration and invasion of cervical cancer cells by inhibiting the expression of MTK-1.
Objective To explore the role if erb-b2 receptor tyrosine kinase 3(ERBB3) in regulating the migration and invasion of cancer cells, and its role in the occurrence and progression of cervical cancer. Methods Totally 25 patients with cervical squamous cell carcinoma and 25 patients with cervical adenocarcinoma were enrolled who were treated in our center from April 2014 to December 2016. Expression level of ERBB3 in patients with cervical squamous cell carcinoma and cervical adenocarcinoma was detected by qRT-PCR. After ERBB3 siRNA silencing on cervical squamous cell, Transwell experiment was used to analyze the capability of migration and invasion, and CCK-8 kits was used to analyze the capability of proliferation, and Western blotting was used to detect S-methyl-5-thioribose kinase 1(MTK-1) protein level. Results qRT-PCR results showed that the expression levels of ERBB3 decreased in cervical squamous cell carcinoma and cervical adenocarcinoma were lower than those in the normal cervical cell lines(P=0.00, P=0.00). The expression level of HPV infection to ERBB3 protein in cervical squamous cell carcinoma cell lines was 10.30±0.07(HPV positive), 7.99 ±0.75(HPV negative) and normal cervical cell lines were 3.02±0.47(HPV positive) and 3.16±0.22(HPV negative), with no statistical significance(P>0.05). After ERBB3 si RNA silencing, the capabilities of SiHa and C33 A in cervical squamous cell carcinoma in proliferation(0.20±0.10, 0.16±0.08), migration(64.87±0.26, 55.02±0.06), and invasion(49.99±0.06, 62.80±0.19) were significantly lower than those in normal cervical cell lines(P<0.05). In addition, the expression level of MTK-1 protein was also significantly reduced after MTK-1 siRNA silencing. Conclusion Down-regulation of ERBB3 can decrease the proliferation, migration and invasion of cervical cancer cells by inhibiting the expression of MTK-1.
引文
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[18]CastellsaguéX.Natural history and epidemiology of HPV infection and cervical cancer[J].Gynecol Oncol,2008,110(3):S4-S7.DOI:10.1016/j.ygyno.2008.07.045.
[19]Schiffman M,Wentzensen N.Human papillomavirus infection and the multistage carcinogenesis of cervical cancer[J].Cancer Epidemiol Biomarkers Prev,2013,22(4):553-560.DOI:10.1158/1055-9965.EPI-12-1406.
[20]Sollome JJ,Thavathiru E,Camenisch TD,et al.HER2/HER3regulates extracellular acidification and cell migration through MTK1(MEKK4)[J].Cell Signal,2014,26(1):70-82.DOI:10.1016/j.cellsig.2013.08.043.
[2]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.DOI:10.3322/caac.21338.
[3]Yang PM,Chou CJ,Tseng SH,et al.Bioinformatics and in vitro experimental analyses identify the selective therapeutic potential of interferon gamma and apigenin against cervical squamous cell carcinoma and adenocarcinoma[J].Oncotarget,2017,8(28):46145-46162.DOI:10.18632/oncotarget.17574.
[4]Zur Hausen H.Papillomaviruses and cancer:from basic studies to clinical application[J].Nat Rev Cancer,2002,2(5):342-350.DOI:10.1038/nrc798.
[5]Schiffman M,Castle PE,Jeronimo J,et al.Human papillomavirus and cervical cancer[J].Lancet,2007,370(9590):890-907.DOI:10.1016/S0140-6736(07)61416-0.
[6]Burd EM.Human papillomavirus and cervical cancer[J].Clin Microbiol Rev,2003,16(1):1-17.
[7]Hildesheim A,Gonzalez P,Kreimer AR,et al.Impact of human papillomavirus(HPV)16 and 18 vaccination on prevalent infections and rates of cervical lesions after excisional treatment[J].Am J Obstet Gynecol,2016,215(2):212.e1-212.e15.DOI:10.1016/j.ajog.2016.02.021.
[8]Galic V,Herzog TJ,Lewin SN,et al.Prognostic significance of adenocarcinoma histology in women with cervical cancer[J].Gynecol Oncol,2012,125(2):287-291.DOI:10.1016/j.ygyno.2012.01.012.
[9]Cancer Genome Atlas Research Network.Integrated genomic and molecular characterization of cervical cancer[J].Nature,2017,543(7645):378-384.DOI:10.1038/nature21386.
[10]Camacho-Leal MP,Sciortino M,Cabodi S.Erb B2 receptor in breast cancer:implications in cancer cell mgration,invasion and resistance to targeted therapy[M].London:Breast Cancer-From Biology to Medicine.In Tech,2017.
[11]Shi H,Gong H,Cao K,et al.Nrdp1-mediated Erb B3 degradation inhibits glioma cell migration and invasion by reducing cytoplasmic localization of p27Kip1[J].J Neurooncol,2015,124(3):357-364.DOI:10.1007/s11060-015-1851-9.
[12]Roskoski R Jr.The Erb B/HER family of protein-tyrosine kinases and cancer[J].Pharmacol Res,2014,79:34-74.DOI:10.1016/j.phrs.2013.11.002.
[13]Yan X,Chen X,Liang H,et al.mi R-143 and mi R-145 synergistically regulate ERBB3 to suppress cell proliferation and invasion in breast cancer[J].Mol Cancer,2014,13(1):220.DOI:10.1186/1476-4598-13-220.
[14]Michael N,Hopkins M,Jura N.Mechanism of PI3K activation by the HER3/Erb B3 receptor[J].Cancer Res,2016(76):4590.DOI:10.1158/1538-7445.
[15]Lyu H,Huang J,Edgerton SM,et al.Role of Erb B3 in tumorigenesis and drug resistance in Erb B2-driven breast cancer[J].Clin Cancer Res,2015(21):B20.DOI:10.1158/1557-3265.
[16]Ma J,Lyu H,Huang J,et al.Targeting of erb B3 receptor to overcome resistance in cancer treatment[J].Mol Cancer,2014,13(1):105.DOI:10.1186/1476-4598-13-105.
[17]Davies S,Holmes A,Lomo L,et al.High incidence of Erb B3,Erb B4and MET expression in ovarian cancer[J].Int J Gynecol Pathol,2014,33(4):402.DOI:10.1097/PGP.0000000000000081.
[18]CastellsaguéX.Natural history and epidemiology of HPV infection and cervical cancer[J].Gynecol Oncol,2008,110(3):S4-S7.DOI:10.1016/j.ygyno.2008.07.045.
[19]Schiffman M,Wentzensen N.Human papillomavirus infection and the multistage carcinogenesis of cervical cancer[J].Cancer Epidemiol Biomarkers Prev,2013,22(4):553-560.DOI:10.1158/1055-9965.EPI-12-1406.
[20]Sollome JJ,Thavathiru E,Camenisch TD,et al.HER2/HER3regulates extracellular acidification and cell migration through MTK1(MEKK4)[J].Cell Signal,2014,26(1):70-82.DOI:10.1016/j.cellsig.2013.08.043.