REV3基因对结肠癌细胞遗传信息表达的影响
收稿日期: 2009-10-28
修回日期: 2010-01-07
网络出版日期: 2010-05-05
基金资助
国家自然科学基金项目(编号: 30560132), 教育部“春晖计划”项目(编号: Z2006-1-75002)和宁夏自然科学基金项目(编号: NZ0885)资助
Roles of REV3 in proliferation and genomic stability of colon carci-noma cells
Received date: 2009-10-28
Revised date: 2010-01-07
Online published: 2010-05-05
利用RNA干扰技术降低REV3基因在人类结肠癌细胞(SW480)中的表达, 以荧光实时定量PCR检测REV3表达量的降低情况, 选择低表达效率具有统计学意义的细胞作为实验组细胞。运用细胞生长曲线、MTT、微核和姐妹染色单体交换等方法, 对实验组和对照组细胞进行细胞生长周期、增殖变化情况和遗传信息表达等指标的检测。结果显示: REV3低表达的结肠癌实验组细胞在细胞增殖以及细胞的微核和姐妹染色单体交换等遗传信息表达均明显低于结肠癌对照组细胞, 实验结果具有统计学意义(P＜0.05); 结肠癌的两对照组间(阴性和空白)的结果虽然有一定的差异, 但没有统计学意义。研究结果提示, REV3低表达时, 可能对结肠癌细胞(SW480)的生长与增殖产生影响, 并对微核和姐妹染色单体交换等遗传不稳定现象的产生有一定的抑制作用。
关键词： REV3基因; RNA干扰; 结肠癌细胞(SW480); 荧光实时定量PCR; 遗传不稳定
隋御，李元杰，金彩霞，徐方 . REV3基因对结肠癌细胞遗传信息表达的影响[J]. 遗传, 2010 , 32(5) : 467 -472 . DOI: 10.3724/SP.J.1005.2010.00467
To investigate the roles of human REV3 gene in proliferation and genomic stability, we experimentally suppressed the REV3 expression in human colon cancer cells (SW480) by the interference RNA technology (RNAi) as monitored by real-time RT-PCR. Compared to untreated or mock-treated cells, ablation of REV3 significantly reduced cell growth rate, micronucleus formation and the frequency of sister chromatid exchange. Whereas the differences between untreated and mock-treated controls were insignificant. Indicators of cell cycle, proliferation and the expression of genetic information in cells of case group, which displayed lower-level expression of REV3, were extremely lower than the control group, and the difference was significant (P<0.05). Differences in the two control groups were not significant. This suggested that the reduced expression of REV3 may affect the growth and proliferation of colon cancer cells (SW480), and, to some extent, contribute to suppression of the genetic instability occurred in micronuclei and sister chromatids. Based on the results from this study, REV3 plays an important role in different cellular growth periods and physiological conditions, and its differential expression directly affects the development of human colon cancer cells.
[1] 郑树. 我国结直肠癌研究现状. 实用肿瘤杂志, 2006, 24(11): 97-99.
[2] 王媛媛, 蔡振利. 微阵列技术在结肠癌研究中的应用. 国际内科学杂志, 2008, 35(3): 167-169.
[3] Xiao W, Lechler T, Chow BL, Fontanie T, Agustus M, Carter KC, Wei YF. Identification, chromosomal mapping and tissue-specific expression of hREV3 encoding a puta-tive human DNA polymerase zeta. Carcinogenesis, 1998, 19(5): 945-949.
[4] Wang ZG. DNA damage-induced mutagenesis: A novel target for cancer prevention. Molecular Interventions, 2001, 1(5): 269-281.
[5] Yuan B, Xu Y, Woo JH, Wang YY, Bae YK, Yoon DS, Wersto RP, Tully E, Wilsbach K, Gabrielson E. Increased expression of mitotic check point genes in breast cancer cells with chromosomal instability. Clin Cancer Res, 2006, 12(2): 405-410.
[6] Livak KJ, Schmittgen TD. Analysis of relative gene ex-pression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) method. Methods, 2001, 25(4): 402-408.
[7] 司徒镇强, 吴军正. 细胞培养. 西安: 世界图书出版社, 1996, 11-32.
[8] 马玉彦, 王波, 鄢德友, 刘理, 李殿俊, 林英姬, 娄阁, 王惠增, 陈国林. MTT法检测人实体瘤化疗敏感性的临床应用研究. 实用肿瘤杂志, 1996, 10(4): 9-10.
[9] Kozmin SG, Pavlov YI, Kunkel TA, Sage E. Roles of Saccharomyces cerevisiae DNA polymerases Poleta and Pol zeta in response to irradiation by simulated sunlight. Nucleic Acids Res, 2003, 31(15): 4541-4552.
[10] 徐方, 余应年, 胡文蔚. 表达反义hREV3基因片段的真核细胞表达质粒的构建. 中国病理生理杂志, 1999, 15(8): 703-706.
[11] 徐方, 余应年, 宋韬. hREV3基因表达被反义阻断的人胚肾细胞系的建立及其某些生物学特性观察. 中国病理生理杂志, 2000, 16(4): 293-297.
[12] 金彩霞, 徐方, 余应年. 反义阻断REV3基因表达对细胞自发突变和MNNG诱导突变的影响. 宁夏医学院学报, 2002, 24(5): 313-315.
[13] Peter EM, Gibbs W, Mcgregor G, Maher VM, Nisson P, Lawrence CW. A human homology of the Saccharomyces cerevisice, REV3 gene, which encodes the catalytic sub-unit of DNA polymerase. Proc Natl Acad Sci USA, 1998, 95(12): 6876-6880.
[14] Bemark M, Khamlichi AA, Davies SL, Neuberger MS. Disruption of mouse polymerase ζ(Rev3) leads to embry-onic lethality and impairs blastocyst development in vitro. Curr Biol, 2000,10(19): 1213-1216.
[15] 徐方, 李元杰. hREV3 基因对细胞增殖周期的影响. 遗传, 2008, 30(8): 1003-1007.
[16] Esposito G, Godindagger I, Klein U, Yaspo ML, Cumano A, Rajewsky K. Disruption of the Rev3-encoded catalytic subunit of polymerase zeta in mice results in early em-bryonic lethality. Curr Biol, 2000, 10(19): 1221-1224.
[17] Wei QY, Li L, Chen DJ. DNA repair genetic in stability and cancer. Chapter 9. DNA Damage Tolerance and Tran-slesion Synthesis. World Scientific, 2006, 233-266.
[18] Wittschieben JP, Reshmi SC, Gollin SM, Wood RD. Loss of DNA polymerase ζ causes chromosomal instability in mammalian cells. Cancer Res, 2006, 66(1): 134-142.
[19] Brondello JM, Pillaire MJ, Rodriguez C, Gourraud PA, Selves J, Cazaux C, Piette J. Novel evidences for a tumor suppressor role of Rev3, the catalytic subunit of Pol zeta. Oncogene, 2008, 27(47): 6093-6101.
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