Suppression of Mcl-1 via RNA interference sensitizes human hepatocellular carcinoma cells towards apoptosis induction.
Suppression of Mcl-1 via RNA interference sensitizes human hepatocellular carcinoma cells towards apoptosis induction.
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DOI：
10.1186/1471-2407-6-232
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发表时间：
2006-10-02
期刊：
影响因子：
3.8
通讯作者：
Galle PR
中科院分区：
文献类型：
--
作者：
Schulze-Bergkamen H;Fleischer B;Schuchmann M;Weber A;Weinmann A;Krammer PH;Galle PR
肝细胞癌是世界范围内最常见的癌症之一，也是癌症相关死亡的主要原因。肝细胞癌对目前可用的化疗药物高度耐药。细胞凋亡信号的缺陷导致了这种抵抗。髓样细胞白血病-1(Mcl-1)是Bcl2蛋白家族中的一个抗凋亡成员，它干扰线粒体的激活。在先前的研究中，我们已经证明Mcl-1在人肝细胞癌组织中高表达。在本研究中，我们通过RNA干扰技术调控Mcl-1蛋白在肝癌细胞中的表达，并分析其对体外培养的肝癌细胞凋亡敏感性的影响。RNA干扰是通过转染siRNA来特异性地下调肝癌细胞中Mcl-1的表达。用实时定量聚合酶链式反应和Western印迹法检测MCL-1的表达。用流式细胞仪和荧光分析分别检测化疗药物和不同靶向治疗后细胞凋亡和caspase活性的变化。在这里，我们证明了表达Mcl-1的肝癌细胞株对一组化疗药物的治疗表现出低敏感性。然而，用蒽环素衍生物表阿霉素处理后，肝癌细胞的凋亡率相对较高。抑制PI3K能显著增加化疗诱导的细胞凋亡。RNA干扰可有效下调肝癌细胞中Mcl-1的表达。MCL-1下调可使肝癌细胞对不同化疗药物增敏。敏化伴随着caspase-3和-9的深刻激活。此外，Mcl-1的下调也增加了PI3K抑制剂治疗后的凋亡率，在较低程度上也增加了mTOR、Raf I和VEGF/PDGF激酶抑制剂治疗后的凋亡率。Mcl-1基因敲除对TRAIL诱导的细胞凋亡无明显反应。此外，MCL-1基因敲除可有效增强肝癌细胞对联合治疗的敏感性：MCL-1基因敲除可显著增强肝癌细胞对化疗结合PI3K抑制的凋亡敏感性。我们的数据表明，通过RNA干扰特异性下调Mcl-1是一种很有前途的方法，可以提高肝癌细胞对化疗和分子靶向治疗的敏感性。
Hepatocelluar carcinoma (HCC) is one of the most common cancers worldwide and a major cause of cancer-related mortality. HCC is highly resistant to currently available chemotherapeutic drugs. Defects in apoptosis signaling contribute to this resistance. Myeloid cell leukemia-1 (Mcl-1) is an anti-apoptotic member of the Bcl-2 protein family which interferes with mitochondrial activation. In a previous study we have shown that Mcl-1 is highly expressed in tissues of human HCC. In this study, we manipulated expression of the Mcl-1 protein in HCC cells by RNA interference and analyzed its impact on apoptosis sensitivity of HCC cells in vitro. RNA interference was performed by transfecting siRNA to specifically knock down Mcl-1 expression in HCC cells. Mcl-1 expression was measured by quantitative real-time PCR and Western blot. Induction of apoptosis and caspase activity after treatment with chemotherapeutic drugs and different targeted therapies were measured by flow cytometry and fluorometric analysis, respectively. Here we demonstrate that Mcl-1 expressing HCC cell lines show low sensitivity towards treatment with a panel of chemotherapeutic drugs. However, treatment with the anthracycline derivative epirubicin resulted in comparatively high apoptosis rates in HCC cells. Inhibition of the kinase PI3K significantly increased apoptosis induction by chemotherapy. RNA interference efficiently downregulated Mcl-1 expression in HCC cells. Mcl-1 downregulation sensitized HCC cells to different chemotherapeutic agents. Sensitization was accompanied by profound activation of caspase-3 and -9. In addition, Mcl-1 downregulation also increased apoptosis rates after treatment with PI3K inhibitors and, to a lower extent, after treatment with mTOR, Raf I and VEGF/PDGF kinase inhibitors. TRAIL-induced apoptosis did not markedly respond to Mcl-1 knockdown. Additionally, knockdown of Mcl-1 efficiently enhanced apoptosis sensitivity towards combined treatment modalities: Mcl-1 knockdown significantly augmented apoptosis sensitivity of HCC cells towards chemotherapy combined with PI3K inhibition. Our data suggest that specific downregulation of Mcl-1 by RNA interference is a promising approach to sensitize HCC cells towards chemotherapy and molecularly targeted therapies.
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影响因子：
20.3
作者：
Derenne, S;Monia, B;Amiot, M
通讯作者：
Amiot, M
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12.4
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