婆罗双树样基因2干扰对宫颈癌HeLa细胞增殖和凋亡的影响

doi:10.3969/j.issn.1004-616x.2016.05.005

癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (5): 359-363,368.doi: 10.3969/j.issn.1004-616x.2016.05.005

婆罗双树样基因2干扰对宫颈癌HeLa细胞增殖和凋亡的影响

张雪杉, 缪芳, 曹晏宁, 张晓姝

滨州医学院免疫学教研室, 山东烟台 264003

收稿日期:2016-03-24 修回日期:2016-05-26 出版日期:2016-09-30 发布日期:2016-09-30
通讯作者: 张晓姝,E-mail:xiaoshu_cz@163.com E-mail:xiaoshu_cz@163.com
作者简介:张雪杉,E-mail:404427485@qq.com
基金资助:
国家自然科学基金资助项目（81001300）

SALL2 interference on proliferation and apoptosis of cervical carcinoma HeLa cells

ZHANG Xueshan, MIAO Fang, CAO Yanning, ZHANG Xiaoshu

Department of Immunology, Binzhou Medical College, Yantai 264003, Shandong, China

Received:2016-03-24 Revised:2016-05-26 Online:2016-09-30 Published:2016-09-30

摘要/Abstract

摘要： 目的：研究婆罗双树样基因2（sal-like gene 2，SALL2）干扰对宫颈癌HeLa细胞增殖和凋亡的影响。方法：设计针对SALL2基因的3条小干扰RNA（siRNA-1、2、3），并设载体组及未转染对照组，使用脂质体转染HeLa细胞。采用逆转录RT-PCR和Western blot检测转染后HeLa细胞中SALL2 mRNA和蛋白的表达以筛选沉默效果最好的siRNA片段。将最适siRNA片段转染HeLa细胞后，采用CCK-8检测细胞增殖率；流式细胞术检测细胞周期和凋亡率。结果：对照组HeLa细胞高表达SALL2。载体组和转染siRNA-3组SALL2的mRNA和蛋白表达与对照组相比，差异均无统计学意义（P均>0.05），而siRNA-1和siRNA-2转染48 h后，HeLa细胞SALL2的mRNA和蛋白表达均降低，差异均有统计学意义（P均<0.05），其中siRNA-1的沉默效率更好，故后续选择siRNA-1作为干扰组。与对照组相比，转染siRNA-1 48和72 h后细胞的增殖率均升高，差异均有统计学意义（P<0.05）；转染48 h后siRNA-1组处于G0/G1期的细胞比例减少，凋亡率明显降低（P均<0.05）。结论：SALL2基因干扰可明显提高HeLa细胞的增殖率，并降低其凋亡率。提示SALL2基因对宫颈癌HeLa细胞的增殖有抑制作用。

关键词: 宫颈癌, SALL2基因, 细胞增殖, 细胞周期, 凋亡

Abstract: OBJECTIVE: To investigate the interference of sal-like 2 (SALL2) gene on proliferation and apoptosis of the cervical carcinoma HeLa cells. METHODS: 3 small interference RNA (siRNA-1,2,3) which targeted SALL2 were structured,and a carrier group and a control group were set. Liposome was used to transfect SALL2-siRNA into HeLa cells. Expression of SALL2 mRNA and protein in the transfected cells were detected by the method of RT-PCR and Western blot by screening the siRNA fragment with the best silencing effect. After the most suitable siRNA fragment was transfected into HeLa cells,CCK-8 was used to detect the cell proliferation rate,and flow cytometry was used to test cell cycle and apoptosis rates. RESULTS: SALL2 was highly expressed in HeLa cells of the control group. Compared with the control group,SALL2 mRNA and protein expression in the carrier group and the siRNA-3 transfection group showed no statistically significant differences (P>0.05),while those in HeLa cells were significantly reduced after transfected with siRNA-1 and siRNA-2 for 48 h (P<0.05). Among them,siRNA-1 showed the best silencing effect,thus siRNA-1 was subsequently selected as the interference group. In contrast with the control group,cell proliferation rate increased significantly after the transfection with siRNA-1 for 48 and 72 h (P<0.05);the proportion of cells in the G0/G1 phase decreased dominantly and apoptosis rate reduced significantly in the siRNA-1 group after transfection for 48 h (P<0.05). CONCLUSION: SALL2 gene interference improved the proliferation rate and reduced the apoptosis rate in HeLa cells,suggesting that SALL2 gene served as a tumor suppressor gene in cervical carcinoma HeLa cells.

Key words: cervical carcinoma, SALL2 gene, cell proliferation, cell cycle, apoptosis

中图分类号:

R392.1

张雪杉, 缪芳, 曹晏宁, 张晓姝. 婆罗双树样基因2干扰对宫颈癌HeLa细胞增殖和凋亡的影响[J]. 癌变·畸变·突变, 2016, 28(5): 359-363,368.

ZHANG Xueshan, MIAO Fang, CAO Yanning, ZHANG Xiaoshu. SALL2 interference on proliferation and apoptosis of cervical carcinoma HeLa cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(5): 359-363,368.

参考文献

[1] De Celis JF,Barrio R.Regulation and function of Spalt proteins during animal development[J].Int J Dev Biol,2009,53(8/9/10):1385-1398.
[2] Sweetman D,Munsterberg A.The vertebrate spalt genes in development and disease[J].Dev Biol,2006,293(2):285-293.
[3] Basson M,Horvitz HR.The caenorhabditis elegans gene sem-4 controls neuronal and mesodermal cell development and encodes a zinc finger protein[J].Genes Dev,1996,10(15):1953-1965.
[4] 巢浩界,张铭,张一鸣,等.SALL4基因在宫颈癌、宫颈上皮内瘤变组织和血浆中表达的研究[J].南京医科大学学报(自然科学版),2014,34(2):188-191.
[5] Sung CK,Li D,Andrews E,et al.Promoter methylation of the SALL2 tumor suppressor gene in ovarian cancers[J].Mol Oncol,2013,7(3):419-427.
[6] Jemal A,Siegel R,Ward E,et al.Cancer statistics,2008[J].CA Cancer J Clin,2008,58(2):71-96.
[7] Ferlay J,Soerjomataram I,Dikshit R,et al.Cancer incidence and mortality worldwide:Sources,methods and major patterns in GLOBOCAN 2012[J].Int J Cancer,2015,136(6):359-386.
[8] 张艳,李宁丽.人类乳头瘤病毒免疫致病机制[J].现代免疫学,2006,26(6):523-526.
[9] Bosch FX,Burchell A,Schiffman M,et al.Epidemiology and natural history of human papillomavirus infections and type-specific implications in cervical neoplasia[J].Vaccine,2008,26(10):1-16.
[10] Naucler P,Ryd W,Tornberg S,et al.Efficacy of HPV DNA testing with cytology triage and/or repeat HPV DNA testing in primary cervical cancer screening[J].J Natl Cancer Inst,2009,101(2):88-99.
[11] Li D,Tian Y,Ma Y,et al.p150(Sal2) is a p53-independent regulator of p21(WAF1/CIP)[J].Mol Cell Biol,2004,24(9):3885-3893.
[12] Sung CK,Yim H,Gu H,et al.The polyoma virus large T binding protein p150 is a transcriptional repressor of c-MYC[J].PLoS One,2012,7(9):e46486.doi:10.1371/journal.pone.0046486.
[13] Li D,Dower K,Ma Y,et al.A tumor host range selection procedure identifies p150(sal2) as a target of polyoma virus large T antigen[J].Proc Natl Acad Sci USA,2001,98(25):14619-14624.
[14] Ma Y,Singer A,Gozman D,et al.Hsal1 is related to kidney and gonad development and is expressed in Wilms tumor[J].Pediatr Nephrol,2001,16(9):701-709.
[15] LI CM,Guo M,Borczuk A,et al.Gene expression in Wilms' tumor mimics the earliest committed stage in the metanephric mesenchymal-epithelial transition[J].Am J Pathol,2002,160(6):2181-2190.
[16] Parroche P,Touka M,Mansour M,et al.Human papillomavirus type 16 E6 inhibits p21(WAF1) transcription independently of p53 by inactivating p150(Sal2)[J].Virology,2011,417(2):443-448.
[17] Chen G,Burger MM.p150 overexpression in gastric carcinoma:the association with p53,apoptosis and cell proliferation[J].Int J Cancer,2004,112(3):393-398.
[18] Nielsen TO,Hsu FD,O'Connell JX,et al.Tissue microarray validation of epidermal growth factor receptor and SALL2 in synovial sarcoma with comparison to tumors of similar histology[J].Am J Pathol,2003,163(4):1449-1456.
[19] Estilo CL,Oc P,Talbot S,et al.Oral tongue cancer gene expression profiling:Identification of novel potential prognosticators by oligonucleotide microarray analysis[J].BMC Cancer,2009,24(7):9-11.
[20] Wu Z,Cheng K,Shi L,et al.Sal-like protein 2 upregulates p16 expression through a proximal promoter element[J].Cancer Sci,2015,106(3):253-261.
[21] Gu H,Li D,Sung CK,et al.DNA-binding and regulatory properties of the transcription factor and putative tumor suppressor p150(Sal2)[J].Biochim Biophys Acta,2011,1809(4/5/6):276-283.
[22] Farkas C,Martins CP,Escobar D,et al.Wild type p53 transcriptionally represses the SALL2 transcription factor under genotoxic stress[J].PLoS One,2013,8(9):e73817.doi:10.1371/journal.pone.0073817.eCollection 2013.

婆罗双树样基因2干扰对宫颈癌HeLa细胞增殖和凋亡的影响

SALL2 interference on proliferation and apoptosis of cervical carcinoma HeLa cells

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