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机构地区:[1]中山大学附属第一医院消化内科,广东广州510080
出 处:《热带医学杂志》2012年第5期514-517,共4页Journal of Tropical Medicine
基 金:国家自然科学基金(81072044)
摘 要:目的观察白介素32(IL-32)在胃癌细胞株中的表达情况,探讨其在胃癌发生、发展中的意义。方法采用逆转录PCR、real-time PCR及Western blot法分别从基因和蛋白水平上检测IL-32在正常胃上皮细胞GES-1以及胃癌细胞株MKN-45、KATO中的表达情况。结果逆转录PCR及real-time PCR结果均显示胃癌细胞IL-32 mRNA表达水平明显高于正常胃上皮细胞,逆转录PCR灰度比值(目的 /内参)为:GES-1(0.34±0.09)、MKN-45(0.79±0.11)、KATO(0.90±0.17),差异有统计学意义(P<0.05)。real-time PCR Ct值显示:MKN-45的IL-32基因表达量是GES-1的(5.34±1.09)倍,KATO是GES-1的(4.07±1.69)倍,差异均有统计学意义(P<0.05)。Western blot结果显示IL-32在蛋白表达水平的结果与前一致,灰度比值(目的 /内参)为:GES-1(0.30±0.10)、MKN-45(0.92±0.32)、KATO(0.86±0.15),差异有统计学意义(P<0.05)。结论IL-32在胃癌细胞株中呈高表达,推测其可能在胃癌的发生、发展中起一定作用。Objective To observe the interleukin-32 expression in gastric cancer cell lines,and to discuss its roles in the development of the gastric cancer.Methods RT-PCR and real time-PCR were used to detect the expression levels of IL-32 mRNA in the human gastric cancer cell lines(MKN-45 and KATO) and the normal gastric epithelial cell GES-1.Western blot was used to detect the expression levels of protein of IL-32.Results Levels of IL-32 mRNA in the human gastric cancer cell lines were significantly higher than that in the normal gastric epithelial cell.The gray-scale ratio(target genes /internal control)were: GES-1(0.34±0.09),MKN-45(0.79±0.11)and KATO(0.90±0.17)(P0.05).The Ct values of real-time PCR showed that the expression level of IL-32 mRNA in MKN-45 was(5.34±1.09)-fold higher than that in GES-1,and KATO were(4.07±1.69)-fold higher.The gray-scale ratio of Western blot were: GES-1(0.30±0.10),MKN-45(0.92±0.32)and KATO(0.86±0.15).It showed that IL-32 protein levels in the human gastric cancer cell lines were significantly higher than that in the normal gastric epithelial cell(P0.05).Conclusion The results showed that the expression of IL-32 in human gastric cancer cell lines were significantly higher than that in the normal gastric epithelial cell.It indicated that the high expression of IL-32 was closely related with progress of human gastric cancer.
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