中国癌症杂志 ›› 2017, Vol. 27 ›› Issue (5): 359-367.doi: 10.19401/j.cnki.1007-3639.2017.05.007

• 论著 • 上一篇    下一篇

RORα高表达对二烯丙基二硫抑制人胃癌MGC803细胞上皮-间质转化的影响

夏 红1,苏 坚1,2,赵晓红1,3,刘 芳1,苏 波1,凌 晖1,曾 希1,苏 琦1   

  1. 1. 湖南省胃癌研究中心,湖南省高校肿瘤细胞与分子病理学重点实验室,
    南华大学肿瘤研究所,湖南 衡阳 421001 ;
    2. 南华大学附属第二医院病理科,湖南 衡阳 421001 ;
    3. 海南省妇幼保健院妇产科,海南 海口 570206
  • 出版日期:2017-05-30 发布日期:2017-06-14
  • 通信作者: 苏 琦 E-mail:suqi1945@163.com
  • 基金资助:
    国家自然科学基金(81374013)。

Effect of RORα overexpression on inhibition of epithelial-mesenchymal transformation by DADS in human gastric MGC803 cells

XIA Hong1, SU Jian1,2, ZHAO Xiaohong1,3, LIU Fang1, SU Bo1, LING Hui1, ZHEN Xi1, SU Qi1   

  1. 1. Center for Gastric Cancer Research of Hunan Province, Key Laboratory of Cancer Cellular and Molecular Pathology of Hunan Provincial University, Cancer Research Institute, University of South China, Hengyang 421001, Hunan Province, China; 2. Department of Pathology, the Second Affiliated Hospital, University of South China, Hengyang 421001, Hunan Province, China; 3. Department of Gynaecology and Obstetrics, Hainan Maternal and Child Health Hospital, Haikou 570206, Hainan Province, China
  • Published:2017-05-30 Online:2017-06-14
  • Contact: SU Qi E-mail: suqi1945@163.com

摘要: 背景与目的:二烯丙基二硫(diallyl disulfide,DADS)具有抗肿瘤的作用。该研究在DADS上调人胃癌MGC803细胞RORα的基础上,观察DADS与RORα高表达对MGC803细胞上皮-间质转化(epithelialmesenchymal transformation,EMT)的影响。方法:相差显微镜观察MGC803细胞形态的影响。采用反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)、蛋白[质]印迹法(Western blot)、免疫荧光与免疫组织化学检测EMT相关分子表达。裸鼠实验检测对移植瘤生长的影响。结果:相差显微镜显示,DADS组与RORα高表达组细胞大小较MGC803细胞一致,呈圆形或椭圆形,梭形细胞少见,异型性降低。RORα高表达加DADS后,上述改变更为明显。Western blot检测结果显示,DADS组与RORα高表达组Snail蛋白表达明显下调,DADS+RORα高表达组更明显(P<0.05)。RT-PCR与Western blot检测显示,DADS与RORα高表达可明显下调Vimentin和上调E-cadherin mRNA与蛋白表达,DADS+RORα高表达组更为显著(P<0.05)。免疫荧光显示,DADS组与RORα高表达组细胞Snail与Vimentin表达较对照组明显减弱和E-cadherin表达显著增强,DADS+RORα高表达组更为显著,与Western blot结果一致。裸鼠实验显示,DADS组、RORα高表达组与RORα高表达+DADS组移植瘤较对照组生长减慢(P<0.05),并且,移植瘤体重较对照组明显降低,抑瘤率分别为15.07%、26.55%与49.27%(P<0.05)。免疫组织化学检测结果显示,DADS组、RORα高表达组与RORα高表达+DADS组较对照组的Ki-67、CD34与Vimentin表达均明显降低,E-cadherin表达明显增强。结论:RORα高表达可增强DADS体内外抑制人胃癌细胞EMT的作用。

关键词: 二烯丙基二硫, ROR&alpha, 人胃癌MGC803细胞, 上皮-间质转化, Snail, Vimentin, E-cadherin

Abstract: Background and purpose: Diallyl disulfide (DADS) could inhibit the growth of cancer cells. This study aimed to investigate the effect of DADS and overexpression of RORα on EMT in human gastric cancer MGC803 cells with upregulation of RORα by DADS. Methods: The morphological effect on MGC803 cells was observed by phase-contrast microscope. The correlative molecules with EMT were detected by RT-PCR, Western blot, immunofluorescence and immunohistochemistry. The influence on xenograft tumor growth in nude mice was observed in MGC803 cells. Results: Phase-contrast microscope showed that MGC803 cells were of identical size, round or oval with decreased spindle cells and lower level of heteromorphism in DADS group and RORα/MGC803 group. The above-mentioned alterations were more obvious in RORα/MGC803+DADS group. Western blot exhibited obviously the downregulation of Snail protein in DADS group and RORα/MGC803 group (P<0.05). RT-PCR and Western blot disclosed that the expression of Vimentin was downregulated notably and E-cadherin was upregulated in DADS group, RORα/MGC803 group, and RORα/MGC803+DADS group more obviously (P<0.05). Immunofluorescence revealed that the positive expression of Snail and Vimentin protein was attenuate, while E-cadherin was strengthened in DADS group, RORα/MGC803 group and RORα/MGC803+DADS group compared with MGC803 cells. Moreover, the xenograft tumor growth was markedly decreased, and body weight of transplanted tumor was visibly reduced with the inhibition ratio of 15.07%, 26.55 % and 49.27%, respectively (P<0.05). The positive expression of Ki-67, CD34 and Vimentin were obviously decreased, while the positive expression of E-cadherin was increased. Conclusion: Overexpression of RORα can remarkably enhance inhibition of EMT in MGC803 cells by DADS in vivo and in vivo.

Key words: Diallyl disulfide, RORα, Human gastric cancer MGC803 cells, Epithelial-mesenchymal transformation, Snail, Vimentin, E-cadherin