雄激素受体对前列腺癌IgG蛋白表达及细胞增殖和迁移的作用
发布时间：2018-10-08 11:06
【摘要】：研究背景:在男性泌尿生殖系统中,前列腺癌是常见的恶性肿瘤。随着社会的的发展,人口老龄化及生活习惯等发生改变,包括中国在内的亚洲地区,前列腺癌发病率呈逐年上升趋势,已位居该地区泌尿系肿瘤的第二位。目前研究已经提示前列腺癌发生及发展的病理进程与雄激素受体(AR)密切相关,前列腺癌细胞恶性程度增加,其雄激素受体的表达减少;早期前列腺癌组织中雄激素受体的表达高于晚期,高度及中度分化的前列腺癌组织中雄激素受体的表达高于低分化前列腺癌。研究发现上皮来源的恶性肿瘤细胞可分泌免疫球蛋白G(IgG),并具有促进肿瘤细胞生长、迁移的作用。在前期研究中,我们发现前列腺癌中表达IgG,其表达随着前列腺癌恶性程度增加而增高,提示IgG可能在前列腺癌发病过程中起重要作用。那么,前列腺癌AR与IgG之间是否存在相关性及其作用机制?目前国内外尚未有相关研究报道。我们拟通过干预前列腺癌细胞株中AR的表达,检测IgG表达及前列腺癌细胞增殖、迁移等生物学特性的变化,明确前列腺癌AR与IgG之间关系。研究目的:研究调控前列腺癌细胞中AR对IgG表达的影响,及对前列腺癌细胞增殖、迁移的作用。研究方法:(1)Western blot分别检测雄激素依赖性前列腺癌细胞株LNCap细胞及去势抵抗性前列腺癌细胞株PC-3细胞中AR及IgG蛋白的表达。(2)构建细胞模型AR基因(pCDNA3.1+)转染PC-3构建AR过表达的PC-3-AR细胞,沉默LNCap中AR基因表达构建LNCap-siAR细胞。(3)检测相关指标Western blot检测AR及IgG表达量;Q-PCR检测细胞株中IgG mRNA表达量;四氮甲基唑氮(MTT)、细胞划痕方法检测细胞的增殖及迁移能力。(4)统计学方法采用SPSS20.0统计软件进行数据处理,数据用x± s表示,PC-3,PC-3-V,PC-3-AR 及 LNCap,LNCap-siCon,LNCap-siAR 多组细胞间 IgG mRNA 及蛋白表达量采用单因素方差分析(采用Bonferroni方法),检验水准α= 0.05。研究结果:(1)LNCap细胞与PC-3细胞相比较,AR高表达,IgG低表达(P0.05)。(2)PC-3细胞转染AR基因后,AR表达增高,PC-3-AR细胞较PC-3细胞低表达IgGmRNA、IgG蛋白,PC-3-AR细胞较PC-3细胞增殖、迁移能力减弱(p0.01)。LNCap细胞沉默AR基因后,AR蛋白表达降低,LNCap-siAR细胞较LNCap细胞高表达IgG mRNA及IgG蛋白,LNCap-siAR细胞较LNCap细胞增殖、迁移能力增强(p0.01)。研究结论:调控前列腺癌细胞AR可影响IgG蛋白的表达,并使其增殖、迁移能力发生改变。上调前列腺癌细胞AR的表达可导致IgG低表达,肿瘤细胞增殖、迁移能力减弱,肿瘤恶性程度降低。沉默前列腺癌细胞AR的表达可导致IgG高表达,肿瘤细胞增殖、迁移能力增强,肿瘤恶性程度增加。
[Abstract]:Background: prostate cancer is a common malignant tumor in male genitourinary system. With the development of society, the aging population and living habits have changed. In Asia, including China, the incidence of prostate cancer has been increasing year by year, and has ranked second among the urinary system tumors in this region. The present study has shown that the pathological process of the occurrence and development of prostate cancer is closely related to the androgen receptor (AR). The malignant degree of prostate cancer cell increases and the expression of androgen receptor decreases. The expression of androgen receptor in early prostate cancer was higher than that in advanced stage, and the expression of androgen receptor in highly and moderately differentiated prostate cancer was higher than that in poorly differentiated prostate cancer. It was found that epithelial malignant tumor cells secreted immunoglobulin G (IgG), and promoted the growth and migration of tumor cells. In previous studies, we found that the expression of IgG, in prostate cancer increased with the increase of the malignant degree of prostate cancer, suggesting that IgG may play an important role in the pathogenesis of prostate cancer. Is there a correlation between AR and IgG in prostate cancer and its mechanism? At present, there is no related research report at home and abroad. We intend to investigate the relationship between prostate cancer AR and IgG by interfering with the expression of AR in prostate cancer cell lines and detecting the expression of IgG and the changes of biological characteristics such as proliferation and migration of prostate cancer cells. Aim: to investigate the effects of AR on the expression of IgG and the proliferation and migration of prostate cancer cells. Methods: (1) Western blot was used to detect the expression of AR and IgG in androgen-dependent prostate cancer cell line LNCap and castrated resistant prostate cancer cell line PC-3, respectively. (2) PC-3 was transfected with AR gene (pCDNA3.1) to construct AR overexpression PC-3-AR cells. LNCap-siAR cells were constructed by silencing AR gene expression in LNCap. (3) AR and IgG expression levels were detected by Western blot and IgG mRNA expression in cell lines was detected by Q-PCR. The cell proliferation and migration were detected by (MTT), cell scratch method. (4) the data were processed by SPSS20.0 software. The expression of IgG mRNA and protein in LNCap,LNCap-siCon,LNCap-siAR and PC-3 PC-3-AR cells was expressed by x 卤s. The expression of IgG mRNA and protein was analyzed by single factor ANOVA (Bonferroni method), and the test level was 伪 = 0.05. Results: (1) compared with PC-3 cells, LNCap cells had higher expression of AR than PC-3 cells (P0.05). (2). After transfection of AR gene, the expression of AR in PC-3-AR cells was higher than that in PC-3 cells. The proliferation of PC-3-AR cells was higher than that of PC-3 cells, and the expression of IgGmRNA,IgG protein in PC-3-AR cells was lower than that in PC-3 cells. The expression of AR protein in LNCap-siAR cells was lower than that in LNCap cells (p0.01). The expression of IgG mRNA in LNCap-siAR cells was higher than that in LNCap cells, and the proliferation of LNCap-siAR cells was higher than that of LNCap cells (p0.01), and the migration ability of LNCap-siAR cells was higher than that of LNCap cells (p0.01). Conclusion: the regulation of AR can affect the expression of IgG protein and change its proliferation and migration ability. Upregulation of AR expression in prostate cancer cells resulted in low expression of IgG, decreased proliferation and migration of tumor cells, and decreased tumor malignancy. Silencing the expression of AR in prostate cancer cells resulted in high expression of IgG, increased proliferation and migration of tumor cells, and increased malignancy of tumor cells.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.25
本文编号：2256521
[Abstract]:Background: prostate cancer is a common malignant tumor in male genitourinary system. With the development of society, the aging population and living habits have changed. In Asia, including China, the incidence of prostate cancer has been increasing year by year, and has ranked second among the urinary system tumors in this region. The present study has shown that the pathological process of the occurrence and development of prostate cancer is closely related to the androgen receptor (AR). The malignant degree of prostate cancer cell increases and the expression of androgen receptor decreases. The expression of androgen receptor in early prostate cancer was higher than that in advanced stage, and the expression of androgen receptor in highly and moderately differentiated prostate cancer was higher than that in poorly differentiated prostate cancer. It was found that epithelial malignant tumor cells secreted immunoglobulin G (IgG), and promoted the growth and migration of tumor cells. In previous studies, we found that the expression of IgG, in prostate cancer increased with the increase of the malignant degree of prostate cancer, suggesting that IgG may play an important role in the pathogenesis of prostate cancer. Is there a correlation between AR and IgG in prostate cancer and its mechanism? At present, there is no related research report at home and abroad. We intend to investigate the relationship between prostate cancer AR and IgG by interfering with the expression of AR in prostate cancer cell lines and detecting the expression of IgG and the changes of biological characteristics such as proliferation and migration of prostate cancer cells. Aim: to investigate the effects of AR on the expression of IgG and the proliferation and migration of prostate cancer cells. Methods: (1) Western blot was used to detect the expression of AR and IgG in androgen-dependent prostate cancer cell line LNCap and castrated resistant prostate cancer cell line PC-3, respectively. (2) PC-3 was transfected with AR gene (pCDNA3.1) to construct AR overexpression PC-3-AR cells. LNCap-siAR cells were constructed by silencing AR gene expression in LNCap. (3) AR and IgG expression levels were detected by Western blot and IgG mRNA expression in cell lines was detected by Q-PCR. The cell proliferation and migration were detected by (MTT), cell scratch method. (4) the data were processed by SPSS20.0 software. The expression of IgG mRNA and protein in LNCap,LNCap-siCon,LNCap-siAR and PC-3 PC-3-AR cells was expressed by x 卤s. The expression of IgG mRNA and protein was analyzed by single factor ANOVA (Bonferroni method), and the test level was 伪 = 0.05. Results: (1) compared with PC-3 cells, LNCap cells had higher expression of AR than PC-3 cells (P0.05). (2). After transfection of AR gene, the expression of AR in PC-3-AR cells was higher than that in PC-3 cells. The proliferation of PC-3-AR cells was higher than that of PC-3 cells, and the expression of IgGmRNA,IgG protein in PC-3-AR cells was lower than that in PC-3 cells. The expression of AR protein in LNCap-siAR cells was lower than that in LNCap cells (p0.01). The expression of IgG mRNA in LNCap-siAR cells was higher than that in LNCap cells, and the proliferation of LNCap-siAR cells was higher than that of LNCap cells (p0.01), and the migration ability of LNCap-siAR cells was higher than that of LNCap cells (p0.01). Conclusion: the regulation of AR can affect the expression of IgG protein and change its proliferation and migration ability. Upregulation of AR expression in prostate cancer cells resulted in low expression of IgG, decreased proliferation and migration of tumor cells, and decreased tumor malignancy. Silencing the expression of AR in prostate cancer cells resulted in high expression of IgG, increased proliferation and migration of tumor cells, and increased malignancy of tumor cells.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.25
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