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Effect of carboplatin injection on Bcl-2 protein expression and apoptosis induction in Raji cells.
European Journal of Histochemistry ( IF 2.1 ) Pub Date : 2020-07-09 , DOI: 10.4081/ejh.2020.3134
Peng Lin 1 , Boliang Zhou 2 , Haiying Yao 1 , Ya-Ping Guo 3
Affiliation  

To investigate the effects of carboplatin (CBP) injection on apoptosis induction in the human lymphoma cell line Raji and to explore the underlying mechanism, Raji cells were randomly divided into two treatment groups. Cells in the experimental groups were treated with 15 μM CBP injection, those in the control groups were treated with solvent, and both groups were treated for 24, 48 and 72 h. Cells from each group were collected for subsequent assays. For each group, the relative expression of B-cell lymphoma-2 (Bcl-2) was determined by Western blot (WB), the expression pattern of Bcl-2 was observed by immunocytochemistry (ICC), and cell apoptosis was observed after Hoechst 33342 staining. Real-time PCR detection of the relative expression levels of the Bax and caspase-3 genes in each group of cells were performed. The WB results showed that the relative expression of the Bcl-2 protein significantly decreased 48 and 72 h after treatment in the CBP groups compared with the control groups (P<0.001), and a significant decrease in the expression of this protein was also noted at 48 h vs 24 h, 72 h vs 48 h, and 72 h vs 24 h with extremely significant differences (P<0.001). Moreover, the expression of the Bcl-2 protein decreased as the duration of CBP treatment increased, showing a time-dependent manner. The ICC results were consistent with the WB findings. The expression of the Bcl-2 protein in the CBP treatment group was significantly reduced 48 h and 72 h after treatment compared with the control group (P<0.001). A time-dependent manner was also noted in the expression of this protein, i.e., the expression level decreased gradually at 24, 48, and 72 h after treatment with statistically significant differences (P<0.001). Hoechst 33342 staining showed that the apoptosis rates at the three time points in the treatment groups were significantly higher than those in the control groups (P<0.001), and a time-effect relationship was observed. The apoptosis rate increased over time with a significant difference (P<0.05). The PCR results showed that the Bax and caspase-3 gene expression trend was the same but opposite that of Bcl-2. After treatment for 24 h and 48 h, the gene expression of the medication groups decreased with a very significant difference (P<0.001), and with prolonged action time, the relative expression of the genes in the medication groups showed an upward trend. Comparing 48 h with 72 h and 24 h with 72 h, the gene expression levels also increased, reaching a very significant difference (P<0.001), and there was a certain time dependence. CBP injection significantly reduced the expression of the Bcl-2 protein and induced apoptosis of Raji cells in a time-dependent manner. Moreover, CBP injection can increase the expression levels of the Bax and caspase-3 genes.



中文翻译:


卡铂注射液对 Raji 细胞 Bcl-2 蛋白表达及凋亡诱导的影响。



为了研究卡铂(CBP)注射对人淋巴瘤细胞系Raji诱导凋亡的影响并探讨其潜在机制,将Raji细胞随机分为两个处理组。实验组细胞用15 μM C​​BP注射液处理,对照组细胞用溶剂处理,两组细胞处理时间分别为24、48和72 h。收集每组的细胞用于后续测定。 Western blot(WB)检测各组B细胞淋巴瘤-2(Bcl-2)的相对表达量,免疫细胞化学(ICC)观察Bcl-2的表达模式,Hoechst后观察细胞凋亡情况33342染色。实时荧光定量PCR检测各组细胞中Bax和caspase-3基因的相对表达量。 WB结果显示,与对照组相比,CBP组治疗后48、72 h Bcl-2蛋白相对表达量显着降低(P<0.001),且该蛋白表达量显着降低(P<0.001)。还注意到在 48 小时24 小时、72 小时48 小时以及 72 小时24 小时时具有极其显着的差异 (P<0.001)。此外,Bcl-2蛋白的表达随着CBP治疗持续时间的增加而降低,呈现出时间依赖性。 ICC 结果与 WB 的结果一致。 CBP治疗组治疗后48 h和72 h Bcl-2蛋白表达量较对照组显着降低(P<0.001)。在该蛋白质的表达中还注意到时间依赖性方式,治疗后24、48、72 h表达量逐渐下降,差异有统计学意义(P<0.001)。 Hoechst 33342染色显示,治疗组3个时间点的细胞凋亡率均显着高于对照组(P<0.001),且呈时效关系。细胞凋亡率随时间增加而增加,差异显着(P<0.05)。 PCR结果显示Bax和caspase-3基因表达趋势相同,而Bcl-2基因表达趋势相反。治疗24 h和48 h后,各用药组基因表达量下降,差异极显着(P<0.001),且随着作用时间的延长,各用药组基因相对表达量呈上升趋势。 48 h与72 h、24 h与72 h相比,基因表达量也有所增加,达到极显着差异(P<0.001),且存在一定的时间依赖性。 CBP注射显着降低Bcl-2蛋白的表达并以时间依赖性方式诱导Raji细胞凋亡。此外,CBP注射可以增加Bax和caspase-3基因的表达水平。

更新日期:2020-08-20
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