摘要
目的 探讨维甲酸α受体 (RARα)介导全反式维甲酸 (ATRA)抑制胃癌细胞生长的作用机理。方法 应用Northernblot分析维甲酸受体在胃癌细胞中的表达水平 ,通过脂质体转染方法将senseRARα和antisenseRARα分别转染到细胞中 ,通过MTT方法和软琼脂集落形成实验分析ATRA对转染细胞生长和恶性程度的影响 ,瞬时转染和测定氯霉素乙酰转移酶 (CAT)活性 ,分析维甲酸应答元件 (RARE)的转录活性。结果 ATRA诱导BGC 82 3细胞RARα表达 ,但不能诱导MKN 45细胞RARα表达。在转染细胞中 ,ATRA可以抑制转senseRARα的MKN细胞生长 ,但不能抑制转antisenseRARα的BGC细胞生长。与MKN 45细胞比较 ,ATRA诱导MKN/RARα细胞 βRARE的转录活性较高 ,但与BGC 82 3细胞比较 ,ATRA诱导BGC/aRARα细胞βRARE的转录活性则较低。
Objective To study the mechanism of retinoic acid receptor α(RARα) in mediating growth inhibition of gastric cancer cells by all trans retinoic acid(ATRA).Methods Expression of RAR s was detected by Northern blot. After anti sense RARα or sense RARα had been transfected into gastric cancer cell lines BGC 823 and MKN 45 respectively, the inhibitory effect of ATRA on cell growth in stable clones was analyzed using MTT assay and colony forming assay in soft agar. The transcriptional activation of retinoic acid response element (RARE) was measured by CAT assay.Results ATRA could induce expression of RARα in BGC 823 cells, but not in MKN 45 cells. In stable clones, ATRA could inhibit growth of MKN 45 cells transfected with RARα gene, but could not inhibit that of BGC 823 cells transfected with antisense RARα. Transient transfection and CAT assay showed higher β RAR response element (βRARE) transcriptional activation induced by ATRA in MKN 45 cells transfected with RARα gene compared to parental MKN 45 cells, while lower βRARE transcriptional activation was seen in BGC cells transfected with antisense RARα gene compared to parental BGC 823 cells. Conclusion Sufficient level of RARα is required for growth inhibition of gastric cancer cells by ATRA.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2000年第3期195-197,共3页
Chinese Journal of Oncology
基金
国家自然科学基金!( 3 9880 0 15 )
国家杰出青年科学基金!( 3 982 5 5 0 2 )
关键词
胃肿瘤
维甲酸
维甲酸α受体
基因转染
Stomach neoplasms
Retinoic acid
Retinoic acid receptor α
Gene transfection
