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目的 应用反义基因转染技术探讨肾癌相关新基因GYLZ RCC18与肾癌细胞死亡 (包括坏死及凋亡 )之间的关系 ,并探讨肾癌基因疗法的新途径。 方法 用脂质体包裹的新基因GYLZ RCC18第一编码区起始处的 2 0个碱基的反义和正义寡核苷酸导入肾癌细胞系GRC 1,应用流式细胞技术和伊红排斥实验连续检测GYLZ RCC18反义寡核苷酸对肾癌细胞凋亡、坏死的影响。 结果 导入GYLZ RCC18反义寡核苷酸后 ,可大量杀伤GRC 1细胞并明显促进GRC 1细胞坏死 ,于第 4天达到杀伤高峰约 6 0 % ;同时可连续 8d诱导凋亡峰 ,最高于第 2天达到约 2 2 5 % ,两种作用曲线形状和峰值不完全一致。 结论 GYLZ RCC18是肾癌相关的重要的新的癌基因 ,GYLZ RCC18的表达可通过两种不同的机制即抗癌细胞坏死机制和抗凋亡机制发挥对肾癌细胞的保护作用 ,GYLZ RCC18可能对肾癌细胞生存和耐药至关重要。通过导入反义基因的方法阻断肾癌细胞中GYLZ RCC18的表达 ,可有效地杀伤肾癌细胞。对此新基因的研究将有助于为肾癌的基础研究和临床治疗提供新的思路和途径。
Objective To investigate the relationship between renal cell carcinoma-related gene GYLZ-RCC18 and cell death (including necrosis and apoptosis) using antisense gene transfection and to explore a new way of gene therapy for renal cell carcinoma. Methods The antisense and sense oligonucleotides of 20 bases at the beginning of the first coding region of GYLZ RCC18, a novel gene encapsulated by liposome, were introduced into the renal carcinoma cell line GRC 1 and were applied to flow cytometry and Eosin exclusion The effect of GYLZ RCC18 antisense oligonucleotide on apoptosis and necrosis of renal cell carcinoma was detected continuously by experiments. Results After GYLZ RCC18 antisense oligonucleotide was introduced, GRC 1 cells could be killed in a large amount and GRC 1 cells were significantly necrotic, reaching a peak of 60% at day 4. At the same time, 2 days to reach about 25%, the two curves of the shape and peak are not exactly the same. Conclusions GYLZ RCC18 is an important new oncogene associated with renal cell carcinoma. The expression of GYLZ RCC18 may play a protective role in renal cancer cells through two different mechanisms, namely anti-cancer cell necrosis mechanism and anti-apoptotic mechanism. GYLZ RCC18 may be Kidney cancer cell survival and drug resistance is crucial. The introduction of antisense gene can block the expression of GYLZ RCC18 in renal cancer cells, which can effectively kill renal cancer cells. Research on this new gene will help to provide new ideas and approaches for the basic research and clinical treatment of renal cell carcinoma.