摘要
目的观察外源性Ｒｂ基因对视网膜母细胞瘤（ＲＢ）细胞凋亡的影响。方法建立裸鼠眼玻璃体腔ＲＢ移植瘤模型及构建Ｒｂ基因的逆转录病毒表达载体Ｐｂａｂｅ－Ｒｂ，用脂质体Ｄｏｓｐｅｒ介导法将Ｒｂ基因导入裸鼠ＲＢ移植瘤，采用流式细胞术（ＦＣＭ），光镜、电镜及ＴＵＮＥＬ细胞凋亡原位末端标记法进行ＲＢ细胞凋亡的检测。结果Ｒｂ基因在ＲＢ移植瘤内表达至少持续７天。ＦＣＭ检测发现在Ｒｂ基因转染后第４天，各实验组治疗眼ＲＢ中均可检测到凋亡细胞峰，凋亡细胞百分率高于对照眼；第２０天，各实验组治疗眼细胞凋亡百分率与对照眼比无差异（Ｐ＞０．０５）。透射电镜下可见到典型的早期凋亡细胞及晚期凋亡小体形成。ＴＵＮＥＬ方法可在荧光显微镜下见到发黄绿色荧光的凋亡细胞，光镜下可见到紫红色的被标记的凋亡细胞。结论Ｒｂ基因可诱导ＲＢ移植瘤细胞凋亡，这可能是Ｒｂ基因体内抗癌的又一作用机制。
ObjectiveTo investigate the effect of exogenous Rb gene on apoptosis of vitreous retinoblastoma (RB) transplantation tumor in nude mouse and its mechanism of tumor suppression.MethodsBased on establishing vitreous RB transplantation tumor in nude mouse,constructing retrovirus vector of Rb gene--PBabe-Rb and transfecing it into the RB transplantation model by liposome Dosper,the change of apoptosis of the RB transplantation tumor by flow cytometry (FCM),light microscope,electron microscope and TUNEL method was analysed.ResultsThe transient expression of Rb protein was detected and the duration of its expression was at least for 7 days.At the 4th day after Rb gene transfection,the peak of apoptotic cells was seen in every treated eyes by FCM and the number of apoptotic cells was more than the control eyes.At the 20th day,no statistically significance in the the number of apoptotic cells between the treated eyes and the control eyes (P>0．05).The typical apoptotic cells and apoptotic body were seen under the electron microscope.The apoptotic cells with yellowgreenish fluorescence under the fluoroscope and also the apoptotic cells marked with purple-redish under the light microscope were found by TUNEL method.ConclusionThe induction of apoptosis by Rb gene mighe be another mechanism of its suppressing tumor growth in vivo.
出处
《眼科研究》
CAS
CSCD
北大核心
2002年第1期23-26,共4页
Chinese Ophthalmic Research
基金
国家自然科学基金(39570751)
CMB基金(9509)
中国医学科学院基金资助
