Xu Yugang, Cheng Ming, Mi Lei, Qiu Yunping, Hao Wenli, Li Leping
Department of General Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China AND Department of General Surgery, Tai'an Central Hospital, Tai'an, Shandong, China.
Department of General Surgery, Tai'an Central Hospital, Tai'an, Shandong, China.
Iran J Allergy Asthma Immunol. 2018 Aug 12;17(4):318-325. doi: 10.18502/ijaai.v17i4.91.
Mir-22-3p is associated with many important biological processes, including neuroprotection, tumorigenesis, and various other tumor progressions. Our study aimed to investigate the roles of Mir-22-3p in chemosensitivity of gastrointestinal stromal tumor (GIST-T1) cells to cisplatin and explore its underlying mechanisms. Mir-22-3p high-expressing cell line was established by transfecting GIST-T1 cell line cells with Mir-22-3p mimic. After treatment with cisplatin (10 μM), Cell counting kits-8 (CCK-8) method was used to detect the cell viability. Flow cytometry was applied to measure the degree of cell apoptosis. Scratch wound healing test was used to detect the migration ability of cells. The protein and mRNA levels of the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) pathway-related factors were analyzed by Western blot and qRT-PCR. The mRNA level of Mir-22-3p was increased in transfected GIST-T1 cells compared with that in control cells. The survival rate and Bcl-2/Bax ratio of GIST-T1 cells treated with both Mir-22-3p analogue and cisplatin were significantly decreased, while the apoptosis rate and protein level of caspase-3 were significantly increased (p<0.05). In addition, the mRNA and protein levels of PTENwere significantly increased in cells treated with both Mir-22-3p analogue and cisplatin (p<0.05), while the expression levels of PI3K and Akt were significantly decreased (p<0.05). Mir-22-3p overexpression can increase the chemosensitivity of cisplatin in human gastrointestinal stromal tumor cells by PTEN/PI3K/Akt pathway.
Mir-22-3p与许多重要的生物学过程相关，包括神经保护、肿瘤发生以及各种其他肿瘤进展。我们的研究旨在探讨Mir-22-3p在胃肠道间质瘤（GIST-T1）细胞对顺铂的化学敏感性中的作用，并探索其潜在机制。通过用Mir-22-3p模拟物转染GIST-T1细胞系来建立Mir-22-3p高表达细胞系。用顺铂（10 μM）处理后，采用细胞计数试剂盒-8（CCK-8）法检测细胞活力。应用流式细胞术测量细胞凋亡程度。划痕伤口愈合试验用于检测细胞的迁移能力。通过蛋白质免疫印迹法和定量逆转录-聚合酶链反应（qRT-PCR）分析10号染色体缺失的磷酸酶和张力蛋白同源物（PTEN）/磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）通路相关因子的蛋白质和mRNA水平。与对照细胞相比，转染后的GIST-T1细胞中Mir-22-3p的mRNA水平升高。用Mir-22-3p类似物和顺铂处理的GIST-T1细胞的存活率和Bcl-2/Bax比值显著降低，而半胱天冬酶-3的凋亡率和蛋白质水平显著升高（p<0.05）。此外，用Mir-22-3p类似物和顺铂处理的细胞中PTEN的mRNA和蛋白质水平显著升高（p<0.05），而PI3K和Akt的表达水平显著降低（p<0.05）。Mir-22-3p过表达可通过PTEN/PI3K/Akt通路增加顺铂对人胃肠道间质瘤细胞的化学敏感性。
