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Year : 2014  |  Volume : 10  |  Issue : 1  |  Page : 68-72

Comparison of nucleostemin gene expression in CD133+ and CD133− cell population in colon cancer cell line HT29

1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Department of Biochemistry, School of Pharmacology, Isfahan University, Isfahan, Iran
4 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Correspondence Address:
Noosha Zia-Jahromi
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran
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Source of Support: This work is financially supported by Islamic Azad University, Research and Science Branch, Tehran, Iran, Conflict of Interest: None

DOI: 10.4103/0973-1482.131375

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Background: Nucleostemin has been shown to be essential for proliferation and survival of colon cancer cells. In this study, we evaluate and comparing nucleostemin expression in CD133+ and CD133- colon cancer cell line HT29. Materials and Methods: After preparation and culturing of HT29 cell line, isolation was performed using magnetic cell separation system by CD133 MicroBeads and phycoerythrin conjugated to monoclonal anti-human CD133 monoclonal antibody and analyzed with flow cytometry. For quantitative expression of nucleostemin in HT29, CD133+ and CD133- cells used specific nucleostemin primer and glyceraldehyde 3-phosphate dehydrogenase primer as endogenous control. Results: The results showed the percentage of CD133+ cells in HT29 colon cancer cell line ranged from 36.5% to 41.5%, whereas the percentage of CD133- cell ranged from 58.5% to 63.5%. The expression rate of nucleostemin in HT29, CD133+ and CD133- cells were 1.44 ± 0.78, 1.60 ± 0.70 and 1.00 ± 0.18 (respectively). The comparison of expression rate represents no significant difference in nucleostemin expression in CD133+, CD133- and HT29 colon cancer cells. Conclusion: It is concluded that nucleostemin expression could not be specific in a certain type of cells in colon cancer cell line HT29 and controlling strategies in colon cancer must not be focused on one certain type of colon cancer cells as main expressing nucleostemin gene.

Abstract in Chinese

核干细胞因子基因表达在CD133+和CD133−结肠癌细胞株HT29细胞群中的比较 背景:核干细胞因子被认为是结肠癌细胞增殖和生存的必要因子。本文中,我们评估和比较CD133+和CD133−结肠癌细胞株HT29中核干细胞因子的表达。 材料和方法:首先准备和培育HT29细胞株,通过磁性细胞分选系统分离CD133微粒和与结合于抗人CD133单克隆抗体的藻红蛋白,再用流式细胞计分析。为了定量表达HT29,CD133+和CD133−细胞中的核干细胞因子,使用了特殊的核干细胞因子引物和甘油醛3-磷酸脱氢酶引物作为内源性控制。 结果:结果表明CD133+细胞在HT29结肠癌细胞株的比例在36.5%到41.5%,而CD133−细胞占58.5%到63.5%。核干细胞因子在HT29,CD133+和CD133−细胞中的表达率分别为1.44± 0.78, 1.60± 0.70 和1.00± 0.18。表达率的比较表明核干细胞因子在CD133+、CD133−和HT29结肠癌细胞中的表达无明显差异。 结论:核干细胞因子表达在HT29结肠癌细胞株的某种细胞中不是特异的,结肠癌的控制策略不必集中于某种主要表达核干细胞因子的结肠癌细胞。 关键词:CD133,结肠癌,基因表达,HT29,核干细胞因子

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