Adenovirus-mediated p16/CDKN2 gene transfer suppresses glioma invasion in vitro
Adenovirus-mediated p16/CDKN2 gene transfer suppresses glioma invasion in vitro
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DOI：
10.1038/sj.onc.1201382
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发表时间：
1997-10-23
期刊：
影响因子：
8
通讯作者：
Rao, JS
中科院分区：
文献类型：
--
作者：
Chintala, SK;Fueyo, J;Rao, JS
恶性胶质瘤广泛浸润周围正常脑组织，其弥漫性浸润是成功治疗的最重要障碍之一，最近的研究表明，胶质瘤从低级别到高级别的进展可能取决于新表型的获得和随后添加的遗传缺陷，胶质瘤进展中最常见的异常之一是肿瘤抑制基因p16的失活，这表明p16的缺失与恶性特征的获得相关。与这一假设相一致，我们先前的研究表明，恢复野生型p16活性进入p16无效的恶性胶质瘤细胞改变了它们的表型。为了理解在高级别胶质瘤中p16失活的生物学后果是否包括促进侵袭性，我们使用携带p16/CDKN 2基因cDNA的重组复制缺陷型腺病毒感染p16细胞并在其中表达高水平的p16蛋白。空SNB 19胶质瘤细胞，在两种模型中测试了SNB 19胶质瘤细胞的侵袭：神经胶质瘤细胞通过Matrigel包被的transwell插入物的侵袭和肿瘤细胞球状体在共培养系统中侵入胎鼠脑聚集体，Matrigel侵袭测定显示表达外源性p16的SNB 19细胞表现出显著降低的侵袭。类似地，与腺病毒对照和模拟感染的细胞的侵袭相比，在72小时的时间段内，p16处理的SNB 19细胞对胎鼠脑聚集体的侵袭减少。基质金属蛋白酶-2（MMP-2），一种参与肿瘤细胞侵袭的酶，与亲本SNB 19和载体感染的细胞相比，在表达p16的SNB 19细胞中的活性显著降低。我们的结果表明，在p16缺失的SNB 19胶质瘤细胞中恢复野生型p16活性显著抑制肿瘤细胞侵袭，从而提示p16基因的新功能。
Malignant gliomas extensively infiltrate the surrounding normal brain, and their diffuse invasion is one of the most important barriers to successful therapy, Recent studies indicate that the progression of gliomas from low-grade to high-grade may depend on the acquisition of a new phenotype and the subsequent addition of genetic defects, One of the most frequent abnormalities in the progression of gliomas is the inactivation of tumor-suppressor gene p16, suggesting that loss of p16 is associated with acquisition of malignant characteristics. Consistent with this hypothesis, our previous studies showed that restoring wild-type p16 activity into p16-null malignant glioma cells modified their phenotype, In order to understand whether the biological consequences of p16 inactivation in high-grade gliomas included facilitating invasiveness, we used a recombinant replication-deficient adenovirus carrying the cDNA of the p16/CDKN2 gene to infect and express high levels of p16 protein in p16-null SNB19 glioma cells, Invasion of SNB19 glioma cells was tested into two models: invasion of glioma cells through Matrigel-coated transwell inserts and invasion of tumor-cell spheroids into fetal rat-brain aggregates in a co-culture system, Matrigel invasion assays showed that the SNB19 cells expressing exogenous p16 exhibited significantly reduced invasion, Similarly, invasion of p16-treated SNB19 cells into fetal rat-brain aggregates was reduced during a 72 h time period compared to invasion of the adenovirus-control and mock-infected cells, Expression of matrix metalloproteinase-2 (MMP-2), an enzyme involved in tumor-cell invasion, in SNB19 cells expressing p16 was significantly reduced compared to that of parental SNB19 and vector-infected cells, Our results show that restoring wild-type p16 activity into p16-null SNB19 glioma cells significantly inhibits tumor-cell invasion, thus suggesting a novel function of the p16 gene.