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    CD5分子对套细胞淋巴瘤细胞株Jeko-1化疗敏感性的影响及其机制研究

    Effects of CD5 on the chemotherapy sensitivity of Jeko-1 mantle cell lymphoma cell line and related mechanisms

    • 摘要: 目的 研究CD5分子对套细胞淋巴瘤细胞株Jeko-1的化疗敏感性及其通路的影响。方法 Crispr/cas9法敲除人源套细胞淋巴瘤细胞株Jeko-1(P)CD5基因靶位点,构建CD5阴性Jeko-1(N)细胞。RT-PCR检测Jeko-1(P)与Jeko-1(N)中CD5-E1A与CD5-E1B的表达。MTS法检测柔红霉素、长春新碱对Jeko-1(P)与Jeko-1(N)细胞增殖抑制效应。ELISA法检测Jeko-1(P)与Jeko-1(N)中IL-10表达。Western blot法检测BAY 11-7082处理与未处理的Jeko-1(P)与Jeko-1(N)中核转录因子-κB (NF-κB)信号通路相关p-IKKβ、p-IκB、p-p65蛋白水平。ELISA法检测应用BAY11-7082处理及未处理的Jeko-1(P)与Jeko-1(N)中IL-10的表达情况。结果 Jeko-1(P)细胞表达CD5-E1A、CD5-E1B,Jeko-1(N)细胞不表达CD5-E1A、CD5-E1B,IL-10分泌减少(P<0.05)。柔红霉素及长春新碱处理24 h和48 h后的Jeko-1(N)细胞与Jeko-1(P)细胞相比,细胞增殖抑制率增加(P<0.05)。BAY 11-7082处理Jeko-1(P)与Jeko-1(N)细胞相比,p-IKKβ、p-IκB、p-p65(核內和核外)、IL-10表达水平差异无统计学意义(P>0.05)。与未处理的Jeko-1(P)组相比,BAY11-7082处理的Jeko-1(P)组与Jeko-1(N)组细胞p-IKKβ、p-IκB 、p-p65(核內和核外)、IL-10 表达水平下降,差异有统计学意义(P<0.05)。结论 CD5分子通过NF-κB信号通路上调套细胞淋巴瘤Jeko-1细胞株IL-10的分泌以影响其化疗敏感性。

       

      Abstract: Objective To investigate the effects of CD5 on the chemotherapy sensitivity of Jeko-1 mantle cell lymphoma cell line and underlying mechanisms. Methods CD5 gene in Jeko-1 (P) cells was knocked out by Crispr/cas9 method, obtaining CD5 negative Jeko-1(N) cells. Then, the expression of CD5-E1A and CD5-E1B were measured by RT-PCR and ELISA using Jeko-1(P) and Jeko-1(N) cells. The inhibitory effects of daunorubicin and vincristine on the proliferation of Jeko-1(P) and Jeko-1(N) cells were examined by MTS assay. The levels of IL-10 in Jeko-1(P) and Jeko-1(N) cells were determined by ELISA. The levels of NF-κB signaling pathway related proteins p-IKKβ, p-IκB and p-p65 in Jeko-1(P) and Jeko-1(N) cells with or without exposure to BAY 11-7082 were measured by Western blotting. The levels of IL-10 in Jeko-1(P) and Jeko-1(N) cells with or without exposure to BAY11-7082 were determined by ELISA. Results The expression of CD5-E1A and CD5-E1B were determined in Jeko-1(P) cells rather than Jeko-1(N) cells, with reduced secretion of IL-10 (P<0.05). Compared with Jeko-1(P) cells, Jeko-1(N) cells showed an increased proliferation inhibitory rate after 24 and 48 h of exposure to daunorubicin and vincristine (P<0.05). No significant difference was found as to the levels of p-IKKβ, p-IκB, p-p65 (inside and outside the nucleus) and IL-10 between Jeko-1(N) and Jeko-1(P) cells after- treated with BAY 11-7082 (P>0.05). Compared with Jeko-1(P) cells without exposed to BAY 11-7082, Jeko-1(P) and Jeko-1(N) after exposure to BAY 11-7082 produced remarkably reduced levels of p-IKKβ, p-IκB, p-p65 (inside and outside nucleus) and IL-10 (P<0.05). Conclusions CD5 may affect the chemotherapy sensitivity of Jeko-1 mantle cell lymphoma cell line through up-regulating its secretion of IL-10 via NF-κB signaling pathway.

       

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