CTLA-4 silencing could promote anti-tumor effects in hepatocellular.

IF 2.8 4区 医学 Q2 ONCOLOGY
Amirhossein Mardi, Mahsan Alizadeh, Amir Shahabaddin Abdolalizadeh, Amir Baghbanzadeh, Behzad Baradaran, Ali Aghebaqti-Maleki, Siamak Sandoghchian Shotorbani, Mohammad Movloudi, Leili Aghebati-Maleki
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Abstract

Preclinical and clinical research showed that immune checkpoint blockade provides beneficial effects for many patients with liver cancer. This study aimed to assess the effect of CTLA-4-specific siRNA on the proliferation, cell cycle, migration, and apoptosis of HePG2 cells. Transfection of siRNA was performed by electroporation. The viability of cells was determined through MTT assay. Flow cytometry was performed to investigate the cell cycle and apoptosis rate, and the wound-healing assay was used to determine HepG2 cells migration. The expression levels of CTLA-4, c-Myc, Ki-67, BCL-2, BAX, caspase-9 (CAS9), and MMP-2,9,13 were measured by qRT-PCR. Transfection of specific CTLA-4-siRNA significantly inhibited the expression of the CTLA-4 gene. Also, our results revealed that CTLA-4 silencing diminished the proliferation and migration as well as induced the apoptosis of HePG2 cells. CTLA-4-siRNA transfection induced the cell cycle arrest in G2 phase. Moreover, CTLA-4-siRNA transfection reduced the expression levels of c-Myc, Ki-67, BCL-2, MMP-2,9,13, and elevated the expression levels of BAX and caspase-9. Our results suggest that silencing CTLA-4 through specific siRNA may be a promising strategy for future therapeutic interventions for treating liver cancer.

Abstract Image

沉默CTLA-4可促进肝细胞抗肿瘤作用
临床前和临床研究表明,免疫检查点阻断可为许多肝癌患者带来益处。本研究旨在评估 CTLA-4 特异性 siRNA 对 HePG2 细胞增殖、细胞周期、迁移和凋亡的影响。siRNA 的转染采用电穿孔法。细胞活力通过 MTT 试验测定。流式细胞仪检测细胞周期和凋亡率,伤口愈合试验检测 HepG2 细胞的迁移。qRT-PCR检测了CTLA-4、c-Myc、Ki-67、BCL-2、BAX、caspase-9(CAS9)和MMP-2、9、13的表达水平。转染特异性 CTLA-4-siRNA 能明显抑制 CTLA-4 基因的表达。同时,我们的研究结果表明,沉默 CTLA-4 可减少 HePG2 细胞的增殖和迁移,并诱导其凋亡。CTLA-4-siRNA 转染诱导细胞周期停滞在 G2 期。此外,CTLA-4-siRNA转染降低了c-Myc、Ki-67、BCL-2、MMP-2、9、13的表达水平,并升高了BAX和caspase-9的表达水平。我们的研究结果表明,通过特异性 siRNA 沉默 CTLA-4 可能是未来治疗肝癌的一种有前途的干预策略。
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来源期刊
Medical Oncology
Medical Oncology 医学-肿瘤学
CiteScore
4.20
自引率
2.90%
发文量
259
审稿时长
1.4 months
期刊介绍: Medical Oncology (MO) communicates the results of clinical and experimental research in oncology and hematology, particularly experimental therapeutics within the fields of immunotherapy and chemotherapy. It also provides state-of-the-art reviews on clinical and experimental therapies. Topics covered include immunobiology, pathogenesis, and treatment of malignant tumors.
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