CARD9在乳腺癌组织中的表达及其对乳腺癌细胞生物学行为的影响
发布时间：2019-04-11 15:08
【摘要】：乳腺癌是女性常见的恶性肿瘤,发病率呈上升趋势,且趋向于年轻化。虽然,近年来,乳腺癌的早期诊断、手术方式、放射治疗以及靶向治疗方面已取得很大进展,但预后仍然较差,靶向药物效率低且价格昂贵。研究乳腺癌发生和发展的分子机制,探索新的分子靶标仍然是乳腺癌研究和临床中需要解决的难题之一。胱天蛋白酶募集域蛋白9(caspase recruitment domain containing protein 9,CARD9)是一个重要的衔接蛋白,通过蛋白间的相互作用调节NF-κB等信号通路,在胃B细胞淋巴瘤、肾癌、丙型肝炎病毒相关的肝细胞癌及结肠癌的发生和发展中扮演着重要角色,但其在乳腺癌中的表达及对乳腺癌发生发展的影响尚未见相关报道。本实验利用实时荧光定量PCR和Western-Blotting技术分别检测乳腺癌癌组织、癌旁组织、乳腺癌细胞、正常乳腺细胞中CARD9 mRNA和蛋白的表达情况,分析CARD9表达与乳腺癌患者临床病理特征间的关系。通过WST-1法、平板克隆形成实验、流式细胞术、划痕实验研究CARD9高表达对乳腺癌细胞增殖、凋亡、周期、侵袭转移等生物学行为的影响。分析CARD9表达和NF-κB信号通路激活的相关性,初步探讨CARD9在乳腺癌中的作用机制。结果表明:乳腺癌组织中CARD9 mRNA(p0.01)和蛋白(p=0.012)表达水平显著高于癌旁组织;乳腺癌T-47D细胞(p=0.021)和MCF-7细胞(p=0.014)中CARD9 mRNA表达水平高于正常乳腺细胞Hs 578Bst;乳腺癌ZR-75-1细胞中CARD9 mRNA(p=0.003)和蛋白(p=0.015)表达水平均高于正常乳腺细胞Hs 578Bst。CARD9蛋白表达与肿瘤大小和雌激素受体(ER)表达有关(p值均0.05)。CARD9高表达可促进乳腺癌细胞的增殖(p0.05),但对细胞凋亡、周期、侵袭和转移未见显著性影响。乳腺癌组织中P50(p0.05)和p-IKKα(p0.01)蛋白的表达水平高于癌旁组织,而且乳腺癌组织中CARD9 mRNA表达与P50(r=0.622,p0.001)、P65(r=0.392,p=0.005)、IL-1β(r=0.685,p0.001)和IL-12(r=0.556,p0.001)mRNA表达间呈正相关。本实验的研究结果揭示了CARD9在乳腺癌组织中的表达情况及其与乳腺癌临床病理学的关系。同时发现,CRAD9很可能通过激活NF-κB信号通路促进乳腺癌细胞增殖从而参与乳腺癌发生发展进程。这些研究结果有助于深入了解CRAD9在乳腺癌发生发展及治疗中的潜在价值,为CARD9成为乳腺癌新的治疗靶点提供了理论依据。
[Abstract]:Breast cancer is a common malignant tumor in women. The incidence of breast cancer is increasing and tends to be younger. Although great progress has been made in early diagnosis, surgical methods, radiotherapy and targeted therapy for breast cancer in recent years, the prognosis is still poor, targeting drugs are inefficient and expensive. To study the molecular mechanism of breast cancer development and explore new molecular targets is still one of the difficult problems to be solved in breast cancer research and clinical practice. Caspase-9 (caspase recruitment domain containing protein-9) is an important binding protein that regulates NF- 魏 B signaling pathways in gastric B-cell lymphoma and renal cancer through protein-to-protein interactions. Hepatitis C virus (HCV) plays an important role in the occurrence and development of hepatocellular carcinoma (HCC) and colon cancer. However, the expression of HCV in breast cancer and its effect on the development of breast cancer have not been reported. The expression of CARD9 mRNA and protein in breast cancer tissues, paracancerous tissues, breast cancer cells and normal breast cells were detected by real-time fluorescence quantitative PCR and Western-Blotting, respectively. To analyze the relationship between the expression of CARD9 and the clinicopathological features of breast cancer patients. The effects of high expression of CARD9 on proliferation, apoptosis, cell cycle, invasion and metastasis of breast cancer cells were studied by WST- 1 assay, plate cloning assay, flow cytometry and scratch assay. To analyze the correlation between the expression of CARD9 and the activation of NF- 魏 B signaling pathway, and to explore the mechanism of CARD9 in breast cancer. The results showed that the expression level of CARD9 mRNA (p0.01) and protein (pA0.012) in breast cancer was significantly higher than that in adjacent tissues. The expression of CARD9 mRNA was higher in breast cancer cells than that in normal breast cells (Hs 578Bst). The expression level of CARD9 mRNA was higher in breast cancer cells than that in normal breast cells (Hs 578BST). The expression level of CARD9 mRNA (p < 0. 0003) and protein (p < 0. 015) in breast cancer ZR-75-1 cells was higher than that in normal breast cells (P < 0. 05). The expression of Hs 578Bst.CARD9 protein in breast cancer cells was correlated with tumor size and estrogen receptor (ER) expression (p < 0. 05). .05). High expression of CARD9 can promote the proliferation of breast cancer cells (p0.05), However, there was no significant effect on cell apoptosis, cell cycle, invasion and metastasis. The expression levels of P50 (p0.05) and p-IKK 伪 (p0.01) protein in breast cancer tissues were higher than those in paracancerous tissues, and the expression of CARD9 mRNA in breast cancer tissues was associated with P50 (r = 0.622, p0.001), P65 (r = 0.392, p < 0.005). There was a positive correlation between the expression of IL-1 尾 (r = 0.685, p0.001) and IL-12 (r = 0.556, p0.001) mRNA. The results of this study revealed the expression of CARD9 in breast cancer and its relationship with the clinicopathology of breast cancer. It was also found that CRAD9 may play an important role in the development of breast cancer by activating NF- 魏 B signaling pathway to promote the proliferation of breast cancer cells. These results are helpful to understand the potential value of CRAD9 in the development and treatment of breast cancer and provide a theoretical basis for CARD9 to become a new therapeutic target for breast cancer.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9
本文编号：2456510
[Abstract]:Breast cancer is a common malignant tumor in women. The incidence of breast cancer is increasing and tends to be younger. Although great progress has been made in early diagnosis, surgical methods, radiotherapy and targeted therapy for breast cancer in recent years, the prognosis is still poor, targeting drugs are inefficient and expensive. To study the molecular mechanism of breast cancer development and explore new molecular targets is still one of the difficult problems to be solved in breast cancer research and clinical practice. Caspase-9 (caspase recruitment domain containing protein-9) is an important binding protein that regulates NF- 魏 B signaling pathways in gastric B-cell lymphoma and renal cancer through protein-to-protein interactions. Hepatitis C virus (HCV) plays an important role in the occurrence and development of hepatocellular carcinoma (HCC) and colon cancer. However, the expression of HCV in breast cancer and its effect on the development of breast cancer have not been reported. The expression of CARD9 mRNA and protein in breast cancer tissues, paracancerous tissues, breast cancer cells and normal breast cells were detected by real-time fluorescence quantitative PCR and Western-Blotting, respectively. To analyze the relationship between the expression of CARD9 and the clinicopathological features of breast cancer patients. The effects of high expression of CARD9 on proliferation, apoptosis, cell cycle, invasion and metastasis of breast cancer cells were studied by WST- 1 assay, plate cloning assay, flow cytometry and scratch assay. To analyze the correlation between the expression of CARD9 and the activation of NF- 魏 B signaling pathway, and to explore the mechanism of CARD9 in breast cancer. The results showed that the expression level of CARD9 mRNA (p0.01) and protein (pA0.012) in breast cancer was significantly higher than that in adjacent tissues. The expression of CARD9 mRNA was higher in breast cancer cells than that in normal breast cells (Hs 578Bst). The expression level of CARD9 mRNA was higher in breast cancer cells than that in normal breast cells (Hs 578BST). The expression level of CARD9 mRNA (p < 0. 0003) and protein (p < 0. 015) in breast cancer ZR-75-1 cells was higher than that in normal breast cells (P < 0. 05). The expression of Hs 578Bst.CARD9 protein in breast cancer cells was correlated with tumor size and estrogen receptor (ER) expression (p < 0. 05). .05). High expression of CARD9 can promote the proliferation of breast cancer cells (p0.05), However, there was no significant effect on cell apoptosis, cell cycle, invasion and metastasis. The expression levels of P50 (p0.05) and p-IKK 伪 (p0.01) protein in breast cancer tissues were higher than those in paracancerous tissues, and the expression of CARD9 mRNA in breast cancer tissues was associated with P50 (r = 0.622, p0.001), P65 (r = 0.392, p < 0.005). There was a positive correlation between the expression of IL-1 尾 (r = 0.685, p0.001) and IL-12 (r = 0.556, p0.001) mRNA. The results of this study revealed the expression of CARD9 in breast cancer and its relationship with the clinicopathology of breast cancer. It was also found that CRAD9 may play an important role in the development of breast cancer by activating NF- 魏 B signaling pathway to promote the proliferation of breast cancer cells. These results are helpful to understand the potential value of CRAD9 in the development and treatment of breast cancer and provide a theoretical basis for CARD9 to become a new therapeutic target for breast cancer.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9
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