中文摘要：

目的探索CD5分子对套细胞淋巴瘤化疗药物敏感性的影响及其分子机制。方法Crispr／cas9法敲除人源套细胞淋巴瘤细胞株Jeko-1（P）的CD5基因，构建Jeko—1（N）（CD5negative）细胞；RT—PCR检测Jeko-1（P）与Jeko-1（N）细胞CD5-E1A和CD5-E1B表达；以不同浓度柔红霉素处理2种细胞，MTS法检测细胞增殖抑制效应；流式细胞术检测细胞周期和细胞凋亡；Western blot检测细胞凋亡抑制蛋白B淋巴细胞瘤2蛋白（B—celllymphoma-2，Bcl-2）、人粒细胞白血病1蛋白（myeloidcellleukemia-1，Mcl—1）、B淋巴细胞瘤XL蛋白（B—celllymphoma—xl，Bcl—XL）的表达。结果Jeko-1（N）细胞不表达CD5-E1A、CD5-E1B；柔红霉素处理后，Jeko-1（N）细胞抑制率高于Jeko-1（P）细胞，IC50低于Jeko-1（P）细胞（P〈0．05），G0／G1期细胞比例增加，S期细胞比例减少（P〈0．05），24h、48h细胞凋亡率较Jeko-1（P）增加，Bcl-2、Mcl蛋白表达量较Jeko-1（P）细胞无明显变化，凋亡抑制蛋白Bcl—XL表达量上调。结论CD5基因敲除的Jeko-1细胞株通过调控淋巴瘤细胞阻滞于G0／G1期，以及上调Bcl—XL蛋白的表达增强对柔红霉素的化疗敏感性。

英文摘要：

Objective To investigate the effect of CD5 on daunorubicin chemosensitivity in a mantle cell lymphoma cell line Jeko - 1 and the underlying mechanisms. Methods A CD5 gene knockout Jeko - 1 cell model （ Jeko - 1 （N） ） was established by Crispr/cas9 method. RT- PCR was performed to analyze the expression of CD5 -EIA, CD5 -E1B on CD5 positive cell line Jeko - 1 （ Jeko - 1 （P） ） and CD5 gene knockout cell line Jeko - 1 [ Jeko - 1 （N） ]. MTS assay was carried out to examine cell proliferation inhibition effect and the chemosensitivity to daunorubiein in Jeko - 1 （P） （CD5 positive） and Jeko- l （N） （CD5 negative） mantle cell lymphomas cell lines. Flow cytometry was used to detect cell cycles and apoptosis in Jeko- 1 （P） （CD5 positive） and Jeko- 1 （N） （ CD5 negetive） mantle cell lymphomas cell lines after exposure of daunorubicin. Western blot was employed to assess the expression of inhibitor of apoptosis protein BcL2, BcL - XL, Mcl after the Jeko - 1 （P） （ CD5 positive） and Jeko - 1 （N） （ CD5 negative） cell lines were exposed to daunorubicin. Results CD5 gene knockout mantle cell lymphomas cell line Jeko - 1 （N） couldn＇t express CD5 - E1A, CD5-E1B. After treatment with daunorubicin, cell line Jeko -1 （N） had higher inhibition rate and lower ICs0 compared to cell line Jeko - 1 （P） （ P 〈 O. 05 ）, with a higher cell ratio of G0/G1 phase and lower cell ratio of S phase （ P 〈 0. 05 ）. Cell line Jeko - 1 （N） obtained a higher apoptosis rate than cell line Jeko - 1 （P） at 24 hours and 48 hours af- ter treatment with daunorubicin （P 〈 0.05）. After treatment with daunorubicin, cell line Jeko - 1 （N） had an upregula- tion of Bcl - XL gene （P 〈 0.05）, but levels of BcL2 and McL showed no change （P 〉 0.05 ） between cell lines Jeko - 1 （N） and Jeko - 1 （P）. Conclusions CD5 gene knockout Jeko - 1 cell may enhance daunorubicin chemosensitivity by arresting the cells at G1/G0 phase and upregulating the Be