Directed antisense therapy confirms the role of protein kinase C-α in the tumorigenicity of pancreatic cancer
Directed antisense therapy confirms the role of protein kinase C-α in the tumorigenicity of pancreatic cancer
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DOI：
10.1016/s0039-6060(98)70123-0
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发表时间：
1998-08-01
期刊：
影响因子：
3.8
通讯作者：
Norman, J
中科院分区：
文献类型：
--
作者：
Denham, DW;Franz, MG;Norman, J
背景。蛋白激酶 C (PKC-α) 的 α 异构体的表达水平已被证明与人类胰腺癌的病理分化呈负相关。 方法，我们稳定转染中度分化的胰腺细胞系 (HPAC) 以过表达 PKC-α，并根据原位模型检查与亲本 HPAC 相比的存活率。接下来我们使用PKC-α反义寡核苷酸在体外特异性下调该亚型，并根据原位模型再次检查体内治疗的效果。结果。植入过表达细胞系的动物的死亡率几乎是植入亲本细胞系的动物的两倍（P < .01）。通过 Northern 印迹分析和逆转录酶-聚合酶链反应，用浓度增加的反义寡核苷酸处理下调 PKC-α mRNA。原位植入胰腺癌细胞后，用反义寡核苷酸治疗的动物比仅用载体治疗的动物在统计上存活时间更长（P = .005）。与单独使用媒介物相比，用乱序寡核苷酸治疗还具有生存益处（P < .01）。结论。胰腺癌的致瘤性与体内 PKC-α 的表达直接相关，过表达时生存率降低就证明了这一点。 PKC-α 表达可以在体外直接（反义）和间接（乱序）下调，从而在体内带来显着的生存益处。
Background. The level of expression of the alpha isoform of protein, kinase C (PKC-alpha) has been shown to correlate inversely with the pathologic differentiation of human pancreatic cancers.Methods, We stably transfected a moderately differentiated pancreatic cell line (HPAC) to overexpress PKC-alpha and examined the survival rates compared with parent HPAC according to an orthotopic model. Next we used a PKC-alpha antisense oligonucleotide specifically to down-regulate this isoform in vitro and examine the effect of treatment in vivo again according to the orthotopic model.Results. Animals implanted with the overexpressing cell line had a mortality rate almost twice that of those implanted with the parent cell line (P < .01). Treatment with antisense oligonucleotide in increasing concentrations down-regulated PKC-alpha mRNA by Northern blot analysis and reverse transcriptase-polymerase chain reaction. Animals treated with antisense oligonucleotide after orthotopic implantation of pancreatic cancer cells survived statistically longer than those treated with vehicle alone (P = .005). Treatment with a scrambled oligonucleotide also conferred a survival benefit compared with vehicle alone (P < .01).Conclusions. Tumorigenicity of pancreatic cancer is related directly to PKC-alpha expression in vivo as demonstrated by decreased survival when overexpressed. PKC-alpha expression can be down-regulated directly (antisense) and indirectly (scrambled) in vitro, which subsequently confers a dramatic survival benefit in vivo.