文档介绍：β肾上腺素能受体阻断剂诱导胰腺癌细胞株PC2凋亡的实验研究
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作者:张东,马清涌,沙焕臣,胡恒通,郭坤
【摘要】目的观察β肾上腺素能受体阻断剂是否通过诱导胰腺癌细胞凋亡而产生抑制效应。方法利用特异性β1肾上腺素能受体阻断剂美托洛尔、广谱β肾上腺素能受体阻断剂普萘洛尔、特异性β2肾上腺素能受体阻断剂丁氧胺干预诱导人胰腺癌PC2细胞。RTPCR和Western blot检测PC2细胞β受体的表达。用Hoechst 33342荧光染色,TUNEL染色和Annexin VFITC/PI流式细胞仪观察和分析PC2细胞凋亡情况,用Western blot检测Caspase3,Caspase9和Caspase8的表达。结果 PC2细胞表达β1和β2肾上腺素受体,尤以β2受体的表达占优势。Hoechst 33342荧光染色、TUNEL染色和流式细胞仪分析表明三种受体阻断剂均诱导了PC2细胞的凋亡,TUNEL染色法结果显示丁氧胺诱导的细胞凋亡率[(±)%]强于普萘洛尔[(±)%],而普萘洛尔诱导的细胞凋亡率优于美托洛尔[(±)%]。流式检测结果显示丁氧胺诱导的细胞凋亡率[(±
)%]优于普萘洛尔[(±)%],而普萘洛尔诱导的细胞凋亡率高于美托洛尔[(±)%]。Western blot分析显示三种受体阻断剂诱导了Caspase3和Caspase9活性片段的表达,而Caspase8未受影响。结论β肾上腺素能受体阻断剂可诱导胰腺癌细胞凋亡,主要是通过阻断β2受体发挥内源性凋亡效应。
【关键词】β肾上腺素能受体;胰腺癌细胞;普萘洛尔;凋亡
ABSTRACT: Objective To observe whether βadrenoceptor antagonists can produce inhibitory effects by inducing apoptosis of pancreatic cancer (PC2) cells. Methods The β1adrenoceptor antagonist metoprolol, βadrenoceptor antagonist propranolol and β2adrenoceptor antagonist butoxamine were used to induce apoptosis in PC2 cells. The mRNA and protein expressions of β1 and β2adrenoceptors were analyzed by RTPCR and Western blotting. The apoptosis index was determined by Hoechst 33342 fluorescent staining, TUNEL and Annexin V and FITC/PI FCM assay. The expressions of Caspase3, Caspase9 and Caspase8 were analyzed by Western blotting. Results PC2 cell line expressed mRNA and protein for both of β1 and β2adrenoceptors. The Hoechst staining, TUNEL and FCM assay showed that the three drugs increased the number of apoptotic cells; TUNEL assay indicated that the rate of apoptosis was the highest using butoxamine (
±)% followed by propranolol (±)%, while the least was using metoprolol (±)%. FCM assay showed that the rate of apoptosis was the highest using butoxamine (±)%, followed by propranolol (±)%, while the least was using metoprolol (±)%. βadre