集落刺激因子1受体介导Bax和Bcl-2表达对人鼻咽癌6-10B细胞凋亡的影响
|
摘要
目的:研究过表达集落刺激因子1受体(colony stimulating factor-1 receptor,CSF-1R)对人鼻咽癌(nasopharyngeal carcinoma,NPC)6-10B细胞凋亡的抑制作用及其与Bax和Bcl-2表达之间的关系。方法:体外利用慢病毒构建的CSF-1R过表达载体LV-CSF1R(16957-1)转染到人鼻咽癌6-10B细胞中,实验分转染组和对照组;采用实时荧光定量PCR及Western blotting检测转染后两组细胞中CSF-1R、Bcl-2、Bax的表达情况;CCK-8法检测两组细胞的增殖;流式细胞术检测两组细胞的凋亡情况。结果:转染组的6-10B细胞中其CSF-1 mRNA表达水平明显高于对照组(7.01±0.23 vs 0.09±0.03,P<0.01);Bax mRNA表达水平显著下调(P<0.01),而Bcl-2 mRNA表达水平显著上调(P<0.01)。转染组的6-10B细胞中CSF-1蛋白表达水平明显高于对照组;Bax蛋白表达水平显著下调,而Bcl-2蛋白表达水平稍上调。与对照组相比,转染组6-10B细胞的增殖活力明显提高(P<0.01);凋亡率显著降低[(10.82±0.75)%vs(17.11±0.46)%,P<0.05]。结论:过表达CSF-1R可以通过调节Bax/Bcl-2之间的比例关系来促进鼻咽癌6-10B细胞的恶性增殖,并抑制细胞凋亡。
Objective: To investigate the inhibitory effect of Colony stimulating factor-1 receptor(CSF-1R) on apoptosis of human nasopharyngeal carcinoma 6-10B cells and its relationship with Bax/Bcl-2 expression. Methods: Lentiviral vector over-expressing CSF-1R LV-CSF1 R(16957-1)was constructed and trasfected into 6-10B cells; meanwhile a control group was set; The expression of CSF-1R, Bcl-2 and Bax was detected by Real-time PCR and Western blotting. CCK-8 assay and flow cytometry(FCM) were used to detect the cell proliferation and apoptosis, respectively. Results: The mRNA expression of CSF-1R increased significantly in 6-10B cells of transfection group compared with negative control(NC) group(7.01±0.23 vs 0.09±0.03, P<0.01); The expression of Bax mRNA was significantly down-regulated while the expression of Bcl-2 mRNA was significantly up-regulated in 6-10B cells of transfection group(all P<0.01). The protein expression of CSF-1 in 6-10B cells of transfection group was significantly higher than that of control group, along with the remarkably down-regulated Bax protein expression and slightly up-regulated Bcl-2 protein in transfection group. Compared with NC group, the proliferation vitality of 6-10B cells were extremely increased after transfection(P<0.01); however, the cell apoptosis was significantly reduced([10.82±0.75]% vs [17.11±0.46]%, P<0.05). Conclusion: Over-expression of CSF-1R could promote the malignant growth and inhibit the apoptosis of nasopharyngeal carcinoma 6-10B cells by regulating the ratio of Bax/Bcl-2 expression.
Objective: To investigate the inhibitory effect of Colony stimulating factor-1 receptor(CSF-1R) on apoptosis of human nasopharyngeal carcinoma 6-10B cells and its relationship with Bax/Bcl-2 expression. Methods: Lentiviral vector over-expressing CSF-1R LV-CSF1 R(16957-1)was constructed and trasfected into 6-10B cells; meanwhile a control group was set; The expression of CSF-1R, Bcl-2 and Bax was detected by Real-time PCR and Western blotting. CCK-8 assay and flow cytometry(FCM) were used to detect the cell proliferation and apoptosis, respectively. Results: The mRNA expression of CSF-1R increased significantly in 6-10B cells of transfection group compared with negative control(NC) group(7.01±0.23 vs 0.09±0.03, P<0.01); The expression of Bax mRNA was significantly down-regulated while the expression of Bcl-2 mRNA was significantly up-regulated in 6-10B cells of transfection group(all P<0.01). The protein expression of CSF-1 in 6-10B cells of transfection group was significantly higher than that of control group, along with the remarkably down-regulated Bax protein expression and slightly up-regulated Bcl-2 protein in transfection group. Compared with NC group, the proliferation vitality of 6-10B cells were extremely increased after transfection(P<0.01); however, the cell apoptosis was significantly reduced([10.82±0.75]% vs [17.11±0.46]%, P<0.05). Conclusion: Over-expression of CSF-1R could promote the malignant growth and inhibit the apoptosis of nasopharyngeal carcinoma 6-10B cells by regulating the ratio of Bax/Bcl-2 expression.
引文
[1]万仁强,傅向军,张学辉,等.mi R-21在鼻咽癌中的表达及对鼻咽癌细胞株增殖及凋亡的影响[J].中国耳鼻咽喉颅底外科杂志,2015,21(5):377-382.DOI:10.11798/j.issn.1007-1520.201505007.
[2]GUO Y,ZHU X D.Identification of genes involved in radioresistance of nasopharyngeal carcinoma by integrating gene ontology and protein-protein interaction networks[J].Int J Oncol,2012,40(1):85-92.DOI:10.3892/ijo.2011.1172.
[3]CHITU V,STANLEY E R.Colony-stimulating factor-1 in immunity and inflammation[J].Curr Opin Immunol,2006,18(1):39-48.DOI:10.1016/j.coi.2005.11.006.
[4]KIRMA N,HAMMES L S,LIU Y G,et al.Elevated expression of the oncogene c-fms and its ligand,the macrophage colony-stimulating factor-1,in cervical cancer and the role of transforming growth factor-beta1 in inducing c-fms expression[J].Cancer Res,2007,67(5):1918-1926.DOI:10.1158/0008-5472.
[5]KIRMA N,LUTHRA R,JONES J,et al.Overexpression of the colonystimulating factor(CSF-1)and/or its receptor c-fms in mammary glands of transgenic mice results in hyperplasia and tumor formation[J].Cancer Res,2004,64(12):4162-4170.DOI:10.1158/0008-5472.
[6]HUANG L,XU X M,HAO Y R.Overexpression of CSF-1R in nasopharyngeal carcinoma[J].Rom J Morphol Embryol,2015,56(4):1279-1283.
[7]YANG S,CHEN J,GUO Y,et al.Identification of prognostic biomarkers for response to radiotherapy by DNA microarray in nasopharyngeal carcinoma patients[J].Int J Oncol,2012,40(5):1590-1600.DOI:10.3892/ijo.2012.1341.
[8]YOSHINORI N,CHIKAGE I,KUNIHIRO Y,et al.Apoptosis and expression of Bcl-2 and Bax proteins in invasive ductal carcinoma of the pancreas[J].Pancreas,2001,22(3):230-230.DOI:10.1097/00006676-200104000-00002.
[9]PEYRAUD F,COUSIN S,ITALIANO A.CSF-1R inhibitor development:current clinical status[J].Curr Oncol Rep,2017,19(11):70.DOI:10.1007/s11912-017-0634-1.
[10]ALIGETI S,KIRMA N B,BINKLEY P A,et al.Colony-stimulating factor-1 exerts direct effects on the proliferation and invasiveness of endometrial epithelial cells[J].Fertil Steril,2011,95(8):2464-2466.DOI:10.1016/j.fertnstert.2011.03.026.
[11]PIXLEY F J.Macrophage migration and its regulation by CSF-1[J/OL].Int J Cell Biol,2012:501962[2017-09-11].https://www.hindawi.com/journals/ijcb/2012/501962/.DOI:10.1155/2012/501962.
[12]GAZDAR A F,MINNA J D.Deregulated EGFR signaling during lung cancer progression:mutations,amplicons,and autocrine loops[J].Cancer Prev Res(Phila),2008,1(3):156-160.DOI:10.1158/1940-6207.
[13]GRUESSNER C,GRUESSNER A,GLASER K,et al.Biomarkers and endosalpingiosis in the ovarian and tubal microenvironment of women at high-risk for pelvic serous carcinoma[J].Am J Cancer Res,2014,4(1):61-72.
[14]ESPINOSA I,EDRIS B,LEE C H,et al.CSF1 expression in nongynecological leiomyosarcoma is associated with increased tumor angiogenesis[J].Am J Pathol,2011,179(4):2100-2107.DOI:10.1016/j.ajpath.2011.06.021.
[15]BACHIREDDY P,RAKHRA K,FELSHER D W.Immunology in the clinic review series;focus on cancer:multiple roles for the immune system in oncogene addiction[J].Clin Exp Immunol,2012,167(2):188-194.DOI:10.1111/j.1365-2249.2011.04514.x.
[16]SHIM A H,CHANG R A,CHEN X,et al.Multi-pronged attenuation of macrophage-colony stimulating factor signaling by EpsteinBarr virus BARF1[J].Proc Natl Acad Sci U S A,2012,109(32):12962-12967.DOI:10.1073/pnas.1205309109.
[17]HUANG L,XU X M,HAO Y R.The possible mechanisms of tumor progression via CSF-1/CSF-1R pathway activation[J].Rom J Morphol Embryol,2014,55(2 Suppl):501-506.
[18]贾炜.Bcl-2家族研究进展[J].当代医学.2012,18(3):26-28.DOI:10.3969/j.issn.1009-4393.2012.3.016.
[19]TOMASIN R,GOMES-MARCONDES M C.Oral administration of Aloe vera and honey reduces Walker tumour growth by de-creasing cell proliferation and increasing apoptosis in tumour tissue[J].Phytother Res,2011,25(4):619-623.DOI:10.1002/ptr.3293.
[20]王卫东,陈正.Bcl-2/Bax比率与细胞“命运”[J].中国肿瘤生物治疗杂志,2007,14(4):393-396.DOI:10.3872/j.issn.1007-385X.2007.4.121.
[2]GUO Y,ZHU X D.Identification of genes involved in radioresistance of nasopharyngeal carcinoma by integrating gene ontology and protein-protein interaction networks[J].Int J Oncol,2012,40(1):85-92.DOI:10.3892/ijo.2011.1172.
[3]CHITU V,STANLEY E R.Colony-stimulating factor-1 in immunity and inflammation[J].Curr Opin Immunol,2006,18(1):39-48.DOI:10.1016/j.coi.2005.11.006.
[4]KIRMA N,HAMMES L S,LIU Y G,et al.Elevated expression of the oncogene c-fms and its ligand,the macrophage colony-stimulating factor-1,in cervical cancer and the role of transforming growth factor-beta1 in inducing c-fms expression[J].Cancer Res,2007,67(5):1918-1926.DOI:10.1158/0008-5472.
[5]KIRMA N,LUTHRA R,JONES J,et al.Overexpression of the colonystimulating factor(CSF-1)and/or its receptor c-fms in mammary glands of transgenic mice results in hyperplasia and tumor formation[J].Cancer Res,2004,64(12):4162-4170.DOI:10.1158/0008-5472.
[6]HUANG L,XU X M,HAO Y R.Overexpression of CSF-1R in nasopharyngeal carcinoma[J].Rom J Morphol Embryol,2015,56(4):1279-1283.
[7]YANG S,CHEN J,GUO Y,et al.Identification of prognostic biomarkers for response to radiotherapy by DNA microarray in nasopharyngeal carcinoma patients[J].Int J Oncol,2012,40(5):1590-1600.DOI:10.3892/ijo.2012.1341.
[8]YOSHINORI N,CHIKAGE I,KUNIHIRO Y,et al.Apoptosis and expression of Bcl-2 and Bax proteins in invasive ductal carcinoma of the pancreas[J].Pancreas,2001,22(3):230-230.DOI:10.1097/00006676-200104000-00002.
[9]PEYRAUD F,COUSIN S,ITALIANO A.CSF-1R inhibitor development:current clinical status[J].Curr Oncol Rep,2017,19(11):70.DOI:10.1007/s11912-017-0634-1.
[10]ALIGETI S,KIRMA N B,BINKLEY P A,et al.Colony-stimulating factor-1 exerts direct effects on the proliferation and invasiveness of endometrial epithelial cells[J].Fertil Steril,2011,95(8):2464-2466.DOI:10.1016/j.fertnstert.2011.03.026.
[11]PIXLEY F J.Macrophage migration and its regulation by CSF-1[J/OL].Int J Cell Biol,2012:501962[2017-09-11].https://www.hindawi.com/journals/ijcb/2012/501962/.DOI:10.1155/2012/501962.
[12]GAZDAR A F,MINNA J D.Deregulated EGFR signaling during lung cancer progression:mutations,amplicons,and autocrine loops[J].Cancer Prev Res(Phila),2008,1(3):156-160.DOI:10.1158/1940-6207.
[13]GRUESSNER C,GRUESSNER A,GLASER K,et al.Biomarkers and endosalpingiosis in the ovarian and tubal microenvironment of women at high-risk for pelvic serous carcinoma[J].Am J Cancer Res,2014,4(1):61-72.
[14]ESPINOSA I,EDRIS B,LEE C H,et al.CSF1 expression in nongynecological leiomyosarcoma is associated with increased tumor angiogenesis[J].Am J Pathol,2011,179(4):2100-2107.DOI:10.1016/j.ajpath.2011.06.021.
[15]BACHIREDDY P,RAKHRA K,FELSHER D W.Immunology in the clinic review series;focus on cancer:multiple roles for the immune system in oncogene addiction[J].Clin Exp Immunol,2012,167(2):188-194.DOI:10.1111/j.1365-2249.2011.04514.x.
[16]SHIM A H,CHANG R A,CHEN X,et al.Multi-pronged attenuation of macrophage-colony stimulating factor signaling by EpsteinBarr virus BARF1[J].Proc Natl Acad Sci U S A,2012,109(32):12962-12967.DOI:10.1073/pnas.1205309109.
[17]HUANG L,XU X M,HAO Y R.The possible mechanisms of tumor progression via CSF-1/CSF-1R pathway activation[J].Rom J Morphol Embryol,2014,55(2 Suppl):501-506.
[18]贾炜.Bcl-2家族研究进展[J].当代医学.2012,18(3):26-28.DOI:10.3969/j.issn.1009-4393.2012.3.016.
[19]TOMASIN R,GOMES-MARCONDES M C.Oral administration of Aloe vera and honey reduces Walker tumour growth by de-creasing cell proliferation and increasing apoptosis in tumour tissue[J].Phytother Res,2011,25(4):619-623.DOI:10.1002/ptr.3293.
[20]王卫东,陈正.Bcl-2/Bax比率与细胞“命运”[J].中国肿瘤生物治疗杂志,2007,14(4):393-396.DOI:10.3872/j.issn.1007-385X.2007.4.121.