[大鼠中经单纯疱疹病毒胸苷激酶转染的骨髓基质细胞治疗的抗胶质瘤活性]

[Anti-glioma activity of treatment by bone marrow stromal cells transfected with HSV-tk in the rat].

作者信息

Li Chun-hui, Jiao Bao-hua

机构信息

Department of Neurosurgery, the Second Affiliated Hospital of Hebei Medical University, Shijiazhuang 050000, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2007 Mar;29(3):171-5.

PMID:17649630

Abstract

OBJECTIVE

To study the anti-glioma activity of treatment by bone marrow stromal cells (BMSCs) transfected with AdCMV-tk containing HSV-tk gene in rats.

METHODS

Primary cultured BMSCs were obtained and transfected with HSV-tk (BMSCs/tk) and were injected into contralateral brain of glioma-bearing rats to observe their tropism for glioma cells. RT-PCR was performed to examine the transduct of tk gene after it was transduced into BMSCs. C6 glioma cells were co-cultured with BMSCs transfected with HSV-tk. MTT test was performed to examine its antitumor activity. BMSCs, after being transfected with HSV-tk, were injected into contralateral brain tissue of glioma-bearing rats to show their in vivo antitumor activity. Dynamic MRI was performed to monitor the development of intracranial glioma.

RESULTS

Purified BMSCs were obtained by primary cultured bone marrow cells. After being transfected with HSV-TK, the cells still stably displayed extensive tropism for intracranial glioma and transcripted tk gene. RT-PCR showed that BMSCs/tk were transduced tk gene obviously at 21 days after AdCMV-tk transfection. BMSCs/tk showed a clear bystander effect after being co-cultured with C6 glioma cells in vitro. TUNEL assay showed that BMSCs/tk could obviously show bystander effect and induce apoptosis of glioma cells in vivo with an apoptosis positive ratio of 20.38% +/- 2.57%, showing a statistically significant difference in comparison with BMSCs group (2.56% +/- 0.52%, P = 0.023) and control group (2.74% +/- 0.38%, P = 0.025). Compared with the control group (21.40 +/- 1.63 days), BMSCs/tk transplantation significantly prolonged the survival time of glioma-bearing rats (52.60 +/- 13.11 days, P = 0.000). MRI detection showed that the least volume of intracranial glioma in BMSCs/tk group (8.28 +/- 2.64 mm3), significantly smaller than that in BMSCs group (134.51 +/- 16.37 mm3, P = 0.001) and control group (147.22 +/- 31.05 mm3, P = 0.001). Some of the intracranial gliomaa disappeared after transplantation of BMSCs/tk.

CONCLUSION

BMSCs transfected with AdCMV-tk may become an effective therapy method in the treatment for glioma.

摘要

目的

研究转染含单纯疱疹病毒胸苷激酶（HSV - tk）基因的腺病毒载体（AdCMV - tk）的骨髓基质细胞（BMSCs）对大鼠脑胶质瘤的治疗作用。

方法

获取原代培养的BMSCs，转染HSV - tk基因（BMSCs/tk）后注入荷瘤大鼠对侧脑内，观察其对胶质瘤细胞的趋向性。转导tk基因至BMSCs后，采用RT - PCR检测tk基因的转导情况。将C6胶质瘤细胞与转染HSV - tk的BMSCs共培养，采用MTT法检测其抗肿瘤活性。将转染HSV - tk的BMSCs注入荷瘤大鼠对侧脑组织，观察其体内抗肿瘤活性。采用动态磁共振成像（MRI）监测颅内胶质瘤的发展。

结果

通过原代培养骨髓细胞获得纯化的BMSCs。转染HSV - TK后，细胞仍稳定地对颅内胶质瘤表现出广泛的趋向性，并转录tk基因。RT - PCR显示，AdCMV - tk转染后21天，BMSCs/tk明显转导了tk基因。体外与C6胶质瘤细胞共培养后，BMSCs/tk显示出明显的旁观者效应。TUNEL检测显示，BMSCs/tk在体内可明显发挥旁观者效应，诱导胶质瘤细胞凋亡，凋亡阳性率为20.38%±2.57%，与BMSCs组（2.56%±0.52%，P = 0.023）和对照组（2.74%±0.38%，P = 0.025）相比，差异有统计学意义。与对照组（21.40±1.63天）相比，BMSCs/tk移植显著延长了荷瘤大鼠的生存时间（52.60±13.11天，P = 0.000）。MRI检测显示，BMSCs/tk组颅内胶质瘤体积最小（8.28±2.64 mm³），明显小于BMSCs组（134.51±16.37 mm³，P = 0.001）和对照组（147.22±31.05 mm³，P = 0.001）。BMSCs/tk移植后部分颅内胶质瘤消失。