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沉默该基因通过抑制Akt的磷酸化增强儿童白血病细胞对化疗药物的敏感性。

Silencing the Gene Enhances the Sensitivity of Childhood Leukemia Cells to Chemotherapy Drugs by Suppressing the Phosphorylation of Akt.

作者信息

Liang Xiuling, Xin Xianfang, Qi Dongmei, Fu Chengyan, Ding Mingde

机构信息

Department of Pediatric Internal Medicine, Affiliated Hospital of Taishan Medical University, Tai'an, China.

Department of Gynecology, Affiliated Hospital of Taishan Medical University, Tai'an, China.

出版信息

Yonsei Med J. 2019 Feb;60(2):182-190. doi: 10.3349/ymj.2019.60.2.182.

Abstract

PURPOSE

This study aimed to investigate the effects of on the sensitivity of acute B lymphocytic leukemia cells (Nalm-6 cells) to chemotherapy drugs.

MATERIALS AND METHODS

Children's normal B lymphocytes and Nalm-6 cells were cultured. Nalm-6 cells were transfected with PIK3CA siRNA (siPIK3CA group) or its negative control (PIK3CA-Control group). Normal Nalm-6 cells were named Mock group. Nalm-6 cells transfected by PIK3CA siRNA were treated with Akt inhibitor (siPIK3CA+Akti-1/2 group). mRNA and protein expression was detected by qRT-PCR and Western blot. Proliferation and sensitivity to chemotherapeutic drugs was detected by MTT assay. Cell cycle and apoptosis was explored by low cytometry. Transwell assay was performed to test invasion.

RESULTS

PIK3CA mRNA (=0.008) and protein (=0.006) expression was higher in Nalm-6 cells than that in normal B lymphocytes. Compared with the Mock group and PIK3CA-Control group, Nalm-6 cells of the siPIK3CA group had lower OD495 values (all <0.05) and invasion cell numbers (=0.03 and =0.025), as well as a higher proportion of G0/G1 phase cells (=0.020 and =0.022), percentage of apoptosis (=0.016 and =0.022), and inhibition rate (all <0.05). pAkt expression in the siPIK3CA group (=0.026 and =0.031) and siPIK3CA+Akti-1/2 group (=0.019 and =0.023) was lower than that in the Mock group.

CONCLUSION

PIK3CA silencing inhibited Nalm-6 cell proliferation and invasion, and promoted their apoptosis and sensitivity to chemotherapeutic drugs, potentially through regulation of the PI3K/AKT signaling pathway.

摘要

目的

本研究旨在探讨[未提及内容]对急性B淋巴细胞白血病细胞(Nalm-6细胞)化疗药物敏感性的影响。

材料与方法

培养儿童正常B淋巴细胞和Nalm-6细胞。用PIK3CA siRNA转染Nalm-6细胞(siPIK3CA组)或其阴性对照(PIK3CA-Control组)。正常Nalm-6细胞命名为Mock组。用Akt抑制剂处理经PIK3CA siRNA转染的Nalm-6细胞(siPIK3CA+Akti-1/2组)。通过qRT-PCR和蛋白质印迹法检测mRNA和蛋白质表达。通过MTT法检测增殖和对化疗药物的敏感性。通过流式细胞术探究细胞周期和凋亡。进行Transwell实验检测侵袭。

结果

Nalm-6细胞中PIK3CA mRNA(=0.008)和蛋白质(=0.006)表达高于正常B淋巴细胞。与Mock组和PIK3CA-Control组相比,siPIK3CA组的Nalm-6细胞OD495值较低(均<0.05),侵袭细胞数较少(=0.03和=0.025),G0/G1期细胞比例较高(=0.020和=0.022),凋亡百分比(=0.016和=0.022)以及抑制率较高(均<0.05)。siPIK3CA组(=0.026和=0.031)和siPIK3CA+Akti-1/2组(=0.019和=0.023)的pAkt表达低于Mock组。

结论

沉默PIK3CA抑制Nalm-6细胞增殖和侵袭,并促进其凋亡及对化疗药物的敏感性,可能是通过调节PI3K/AKT信号通路实现的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824b/6342719/7799c10249f1/ymj-60-182-g001.jpg

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