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国家自然科学基金(30600728)

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降低S100A6基因表达对胃癌细胞生物学特性影响的研究被引量:2
2010年
目的:初步探讨S100A6基因对于胃癌细胞生长、增殖状态,细胞周期,凋亡状态等生物学特性及成瘤、浸润转移能力等恶性表型的影响。方法:S100A6基因的RNAi载体通过脂质体介导法转染S100A6基因高表达SGC7901细胞,后经G418压力筛选法获得稳定转染的细胞系,经过RT-PCR法、免疫细胞化学法及Western-bloting法证实。对稳定表达株进行鉴定,而后使用流式细胞仪、生长曲线法、平板克隆形成实验法、细胞迁徙实验等方法分析稳定表达株相关生物学特性与恶性表型的变化,每种检测实验均设立RNAi载体稳定转染细胞组、IMG800空载体稳定转染细胞组和空白SGC7901细胞组。结果:稳定的S100A6基因RNAi细胞系经过RT-PCR法、免疫细胞化学法及Western-bloting法证实,mRNA抑制率可达75%左右,蛋白产物的抑制率达85%左右。与转染IMG800空载体及空白SGC7901胃癌细胞相比转染S100A6 RNAi载体的稳定表达细胞株生长减慢,各时间点细胞计数显著低于对照组(P<0.05);后两组之间亦无显著差异,细胞周期检测显示S100A6 RNA干扰组G0-G1期比例显著高于对照组,而G2-M及S期比例显著低于对照组(P<0.05),其它各期细胞比例均无显著差异。平板克隆形成实验结果显示S100A6 RNA干扰组克隆形成率显著低于对照组(P<0.05);,细胞迁徙实验结果提示S100A6 RNA干扰组穿膜率显著低于对照组(P<0.05)。结论:S100A6基因可能具有促进细胞生长增殖作用,同时可影响细胞周期,增加处于分裂期细胞的比例可能具有促进细胞分裂作用,并可促进胃癌细胞侵袭转移。降低S100A6基因表达可能抑制胃癌细胞的恶性生物学行为。
张林黄海力王孟薇吴本俨吴凯
关键词:胃癌细胞周期RNA干扰
胃癌及其转移灶中S100A6基因的表达被引量:4
2008年
目的分析S100A6在胃癌发展过程中的作用,研究配对胃癌组织以及转移灶中S100A6基因mRNA和蛋白的表达。方法应用实时定量逆转录聚合酶链反应(QRT—PCR)检测20例配对胃癌组织中S100A6mRNA的表达,构建208例胃癌患者配对胃癌组织及转移淋巴结病灶的组织芯片,采用免疫组化方法检测S100A6蛋白的表达,并分析其与临床病理因素以及预后之间的相关性。结果QRT—PCR检测结果显示,20例配对胃癌中,有14例的S100A6mRNA转录水平升高,平均升高倍数为2.25倍。免疫组化检测结果显示,S100A6在正常胃黏膜中的表达阳性率为34.3%,在癌灶中表达阳性率为84.1%,在淋巴结转移灶中表达阳性率为90.9%,癌灶和淋巴结转移灶表达高于正常胃黏膜(P〈0.05)。65.5%的胃癌组织中S100A6表达较对应正常胃黏膜升高。癌灶中S100A6表达与肿瘤大小和侵袭程度相关,肿瘤越大表达越高(P=0.022),侵袭越深表达越高(P=0.000),未发现S100A6表达与其他临床病理因素之间的相关性。S100A6表达与预后无关,但相对于正常胃黏膜,癌灶中S100A6表达升高者的预后比表达下降者差。结论S100A6表达上调是胃癌早期事件,与胃癌的发生、发展相关。
黄海力吴本俨朱旭东尤纬缔王卫华王孟薇
关键词:胃肿瘤基因表达
S100A6 gene have a positive influence on the growth and proliferation of gastric cancer cell MKN45被引量:1
2009年
Objective: S100A6 (a.k.a., calcyclin) is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma (MFH), and carcinomas of the thyroid, breast, and colon. However, little is known about the role S100A6 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of S100A6 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell MKN45. Methods: As an important member of S100 family, S100A6 cDNAwas subcloned into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay respectively. The S100A6 stable expression clones (MKN-S100A6) were detected and compared with their control groups respectively. Results: MKN- S100A6 grew faster than MKN45 and MKN-PC (MKN45 transfected with pcDNA3.1 vector). The cell counts of MKN-SI00A6 in the fifth, sixth and seventh days were significantly more than those of control groups (P 〈 0.05). Cell cycle analysis showed that proportions of MKN-S100A6 in G0-G1 and G2-M were different significantly with those of its control groups respectively (P 〈 0.05). The apoptosis rate of MKN-S100A6 was significantly lower than those of control groups (P 〈 0.05). Results of colony-forming assay showed that the colon formation rate of MKN-S100A6 was higher than those of control groups (P 〈 0.05). Conclusion: S100A6 can promote the growth and proliferation of gastric cancer cells. It can help tumor cell maintain malignant phenotype. In gastric cancer, S100A6 could be thought as a tumor-enhancing gene in some distance, but its role could be complicated.
Lin ZhangYanhong HouNan LiMengwei WangBenyan WuKai Wu
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