Inhibition of hamster mesothelioma tumorigenesis by an antisense expression plasmid to the insulin-like growth factor-1 receptor.
Inhibition of hamster mesothelioma tumorigenesis by an antisense expression plasmid to the insulin-like growth factor-1 receptor.
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DOI：
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发表时间：
1996-09
期刊：
影响因子：
11.2
通讯作者：
H. Pass;D. Mew;M. Carbone;W. Matthews;J. Donington;R. Baserga;C. Walker;M. Resnicoff;S. Steinberg-S.
中科院分区：
文献类型：
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作者：
H. Pass;D. Mew;M. Carbone;W. Matthews;J. Donington;R. Baserga;C. Walker;M. Resnicoff;S. Steinberg-S.
我们评估了反义胰岛素样生长因子（IGF）受体转录物对SV 40诱导的免疫活性仓鼠间皮瘤模型（H9 A）的增殖和致瘤性的影响。用逆转录-PCR和北方分析从H9 A RNA中鉴定IGF-1和IGF-1受体（IGF-1 R）基因的表达。用含有对应于IGF-1 R碱基对1-309的cDNA片段的诱导型表达载体（在热休克启动子HSP 70的转录控制下）以有义或反义方向电穿孔H9 A细胞，以产生相应的克隆A3正义或B 9反义。用PCR分析检测基因组DNA中的表达载体，在溴化乙锭凝胶上作为173-bp的片段。然后在活性（39 ℃）或非活性（34 ℃）条件下体外评价表达载体的作用。在39 ℃下，B 9反义转染子显示出比A3正义转染子显著更少的增殖（P2 < 0.02）。在34 ℃下，A3正义转染细胞和B 9反义转染细胞的细胞生长没有显著差异。在接种10（5）个A3正义或B 9反义转染细胞的仓鼠中评价体内致瘤性。与B 9反义克隆相比，A3正义克隆在体内导致更大数量的肿瘤（P2 = 0.0001）。当从A3正义和B 9反义动物中发展的肿瘤的基因组DNA中分析表达载体时，从表达载体扩增的173-bp片段在正义肿瘤中被鉴定，但在反义B 9或野生型H9 A肿瘤中未被鉴定，表明载体从体内增殖的反义克隆中丢失。IGF-1 R反义转录物对仓鼠间皮瘤的抑制作用在本研究中通过体外和体内生长和致瘤性的降低来证明，这可能对人类间皮瘤的治疗有意义。
We evaluated the effect of antisense insulin-like growth factor (IGF) receptor transcripts on the proliferation and tumorigenicity in an SV40-induced, immunocompetent hamster mesothelioma model (H9A). Expression of IGF-1 and IGF-1 receptor (IGF-1R) genes was identified from H9A RNA using reverse transcription-PCR and Northern analysis. H9A cells were electroporated with inducible expression vectors (under the transcriptional control of heat shock promoter HSP70) containing a cDNA fragment corresponding to base pairs 1-309 of IGF-1R in the sense or antisense orientation to generate the respective clones A3 sense or B9 antisense. The expression vector in genomic DNA was detected with PCR analysis as a 173-bp fragment on ethidium bromide gels. The effects of the expression vectors were then evaluated in vitro under active (at 39 degrees C) or inactive (at 34 degrees C) conditions. At 39 degrees C, the B9 antisense transfectants demonstrated significantly less proliferation than A3 sense transfectants (P2 < 0.02). At 34 degrees C, cell growth of A3 sense- and B9 antisense-transfected cells was not significantly different. In vivo tumorigenicity was evaluated in hamsters inoculated with 10(5) A3 sense- or B9 antisense-transfected cells. The A3 sense clones resulted in greater numbers of tumors in vivo compared to the B9 antisense clone (P2 = 0.0001). When genomic DNA from tumors that developed in A3 sense and B9 antisense animals was analyzed for the expression vectors, a 173-bp fragment amplified from the expression vector was identified in the sense tumors but not in antisense B9 or wild-type H9A tumors, indicating a loss of the vector from the antisense clones that proliferated in vivo. The inhibitory effect of IGF-1R antisense transcripts on hamster mesothelioma demonstrated in this study by decreased growth and tumorigenicity in vitro and in vivo may have implications for the therapy of human mesothelioma.