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肝癌抑制基因-1转运功能区的研究
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作者 周华 刘鸿 +1 位作者 赵新泰 周秀梅 《浙江理工大学学报(自然科学版)》 2011年第2期241-245,282,共6页
肝癌抑制基因-1(hepatocellular carcinoma suppressor 1,HCCS1)是一种潜在的肝癌抑制基因,在细胞内发挥蛋白分拣运输的作用,其抑癌作用有可能与蛋白转运的功能有一定的关系。本研究旨在寻找HCCS1序列中与转运功能相关的序列区域。首先... 肝癌抑制基因-1(hepatocellular carcinoma suppressor 1,HCCS1)是一种潜在的肝癌抑制基因,在细胞内发挥蛋白分拣运输的作用,其抑癌作用有可能与蛋白转运的功能有一定的关系。本研究旨在寻找HCCS1序列中与转运功能相关的序列区域。首先通过亚克隆技术,构建以pEGFP-C2为载体,对不同长度HCCS1-cDNA片段进行亚克隆,将构建的亚克隆转染到HeLa细胞,通过免疫荧光显微镜观察不同长度HCCS1蛋白在细胞内的分布,及其与6-磷酸甘露糖受体(M6PR)的共定位。经过PCR及亚克隆后,目的片段成功插入载体质粒,构建了以pEGFP-C2为载体的含有4个不同长度HCCS1片段的亚克隆。分别是:pEGFP-C2-N1836,pEGFP-C2-N1572,pEGFP-C2-1707和pEGFP-C2-1506,其中pEGFP-C2-N1836和pEGFP-C2-N1572编码的HCCS1蛋白,长度不同均呈颗粒状、极性分布于核周的胞质内,且与M6PR有共定位;而pEGFP-C2-1707片段编码的HCCS1蛋白虽然也呈颗粒状分布于核周,但极性分布消失,且与M6PR共定位消失;pEGFP-C2-1506与pEGFP-C2-1707结果相似。结果显示了HCCS1-cDNA的5’端的129 bp序列为与HCCS1的极性及M6PR共定位相关的区域。 展开更多
关键词 肝癌抑制基因 功能序列 免疫荧光 转运
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肝癌抑制基因-1与转运功能相关的451bp序列区的确定 被引量:1
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作者 孔伟 甘愉 +3 位作者 胡晶莹 朱明洁 赵仰星 赵新泰 《中国癌症杂志》 CAS CSCD 2008年第1期1-5,共5页
背景与目的:肝癌抑制基因-1(HCCS1)是一种潜在的肝癌抑制基因,并且在细胞内蛋白分拣运输中也发挥着重要作用,其抑癌作用有可能是通过其蛋白转运的功能而发挥的,本文以寻找HCCS1序列中与转运相关的最小功能序列区域为目的。方法:通过亚... 背景与目的:肝癌抑制基因-1(HCCS1)是一种潜在的肝癌抑制基因,并且在细胞内蛋白分拣运输中也发挥着重要作用,其抑癌作用有可能是通过其蛋白转运的功能而发挥的,本文以寻找HCCS1序列中与转运相关的最小功能序列区域为目的。方法:通过亚克隆技术,构建以pEGFP-C2为载体的含不同长度HCCS1cDNA片段的亚克隆,将构建的亚克隆转染子宫颈癌HeLa细胞,通过免疫荧光共聚焦显微镜观察不同长度HCCS1蛋白的分布,以及与6-磷酸甘露糖受体(M6PR)的共定位。结果:成功构建了以pEGFP-C2为载体的10个含有不同长度HCCS1片段的亚克隆;HCCS1cDNA从3'端向5'端逐渐缺失的片段中:2100bp片段至778bp片段编码的不同长度HCCS1蛋白呈颗粒状分布于核周的胞质内,其中2100bp片段至1571bp片段编码的不同长度HCCS1蛋白均呈颗粒状、极性分布于核周的胞质内,且与M6PR有共定位;而1120bp片段至778bp片段编码的不同长度HCCS1蛋白虽然也呈颗粒状分布于核周,但极性分布消失,且与M6PR共定位消失;HCCS1cDNA片段678bp片段至339bp片段编码的不同长度HCCS1蛋白呈散点状弥散分布于胞质以及胞核内。结论:确定了HCCS1cDNA1571bp片段的3'端451bp的序列为与HCCS1转运功能相关的最小区域范围。 展开更多
关键词 肝癌抑制基因 功能序列 转运 免疫荧光共聚焦
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Expression and significance of new inhibitor of apoptosis protein survivin in hepatocellular carcinoma 被引量:31
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作者 HongZhu Xiao-PingChen Wan-GuangZhang Shun-FengLuo Bi-XiangZhang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第25期3855-3859,共5页
AM: To investigate expression and significance of inhibitor of apoptosis protein survivin in hepatocellular carcinoma (HCC). METHODS: The expression of survivin and vascular endothelial growth factor (VEGF) was invest... AM: To investigate expression and significance of inhibitor of apoptosis protein survivin in hepatocellular carcinoma (HCC). METHODS: The expression of survivin and vascular endothelial growth factor (VEGF) was investigated in 38 cases of HCC tissues and 38 liver cirrhosis tissues by immunohistochemistry and Western blot. The relationship between the expression of survivin and clinicopathological factors of HCC was analyzed. RESULTS: Survivin protein was detected in 23 (60.5%) of 38 HCCs and 3 (7.9%) of 38 liver cirrhosis tissues. In 23 cases of HCC which expressed survivin, the expression of VEGF was positive in 18 cases and slight positive or negative in 5 cases. While in 15 cases of HCC which did not express survivin, 12 cases did not express or slightly expressed, and 3 cases expressed VEGF. In liver cirrhosis tissues, the expression of VEGF was as follows: 24 cases were negative, 10 cases were weak positive and 4 cases were strong positive. The expression of survivin was coincident with the expression of VEGF in HCC (P<0.01). The expression of survivin in HCC had no relationship with the patients' age, gender, tumor size and differentiation level of HCC, while it was related to the metastasis of HCC. The protein quantitative analysis by Western blot also showed that overexpression of survivin in HCC was closely correlated to the expression of VEGF (P<0.01). Furthermore, stronger expression of survivin and VEGF was also found in patients with metastasis rather than in those with no metastasis (P<0.01). CONCLUSION: Survivin plays a pivotal role in the metastasis of HCC, and it has some correlation with tumorigenesis. The expression of survivin in the primary lesion is very useful as an indicator for metastasis and prognosis of HCC. It could become a new target of gene therapy of HCC. 展开更多
关键词 SURVIVIN Vascular endothelial growth factor Hepatocellular carcinoma
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Melanoma differentiation-associated gene-7, MDA-7/IL-24, selectively induces growth suppression, apoptosis in human hepatocellular carcinoma cell line HepG2 by replication-incompetent adenovirus vector 被引量:15
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作者 Cong-Jun Wang Xin-Bo Xue Ji-Lin Yi Kun Chen Jian-WeiZheng Jian Wang Jian-Ping Zeng Rong-Hua Xu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第11期1774-1779,共6页
AIM: To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma (HCC) cell line HepG2 and normal liver cell line L0... AIM: To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma (HCC) cell line HepG2 and normal liver cell line L02. METHODS: We constructed the recombinant replication-incornpetent Ad.rnda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the rnRNA expressing in cells, by ELISA was used to detect MDA-7/IL-24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.rnda-7 was confirmed by Hoechst staining and flow cytometry assay with Annexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, and cell-cycle and hypodiploidy analyses were performed by flow cytometry. RESULTS: Recombinant replication-defective virus expressing MDA-7/IL-24 was constructed successfully. RTPCR showed that the Ad.rnda-7 could mediate the expression of the exogenous gene MDA-7/IL-24 into HepG2 and L02. The concentration of MDA-7/IL-24 protein in supernatant was 130 pg/mL and 110 pg/mL in Ad.rnda-7-infected L02 and HepG2 ceils, respectively. Ad.mda-7 infection obviously induced apoptosis (from 2.604±0.72% to 33.64±13.2%, P=0.00012) and growth suppression in HepG2 (inhibition ratio IR=68%) and an increase in the percentage of specific cancer cell types at the G2/M phase of the cell cycle (from 6.44% to 32.29%, P〈 0.01), but not in L02 cells.CONCLUSION: These results confirm selectively induction of apoptosis and growth suppression by the mda-7/ IL-24 gene with replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2. 展开更多
关键词 Cancer gene therapy Hepatocellular carcinoma (HCC) APOPTOSIS Growth suppression MDA-7/IL-24
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Decreased expression of Neurensin-2 correlates with poor prognosis in hepatocellular carcinoma 被引量:7
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作者 Hai-Qing Ma Xiao-Ting Liang Jing-Jing Zhao Hui Wang Jian-Cong Sun Yi-Bing Chen Ke Pan Jian-Chuan Xia 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第38期4844-4848,共5页
AIM: To investigate the expression of Neurensin-2 (NRSN2) in hepatocellular carcinoma (HCC) and its prognostic values in predicting survival. METHODS: The expression and prognostic significance of NRSN2 in HCC w... AIM: To investigate the expression of Neurensin-2 (NRSN2) in hepatocellular carcinoma (HCC) and its prognostic values in predicting survival. METHODS: The expression and prognostic significance of NRSN2 in HCC was examined by performing immunohistochemical analysis using a total of 110 HCC clinical tissue samples, and Western blotting analysis to further confirm the result. RESULTS: Decreased NRSN2 expression was shown in 70.9% cases. Loss of NRSN2 expression in HCC was significantly related to tumor size (P = 0.006). Larger tumor size was related to negative expression of NRSN2. Patients showing negative NRSN2 expression had a significantly shorter overall survival than those with positive expression (P = 0.008). Multivariate Cox regression analysis indicated that NRSN2 expression level was an independent factor of survival (P = 0.013). Western blotting analysis further confirmed decreased expression of NRSN2 in tumor tissues compared with non-tumorous tissues. CONCLUSION: Our study indicated that NRSIV2 could be a tumor suppressor gene for HCC and a candidate biomarker for long-term survival in HCC. 展开更多
关键词 Neurensin-2 Hepatocellular carcinoma EXPRESSION Immunohistochemical analysis PROGNOSIS
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