Kishani Farahani Roya, Nazemalhosseini Mojarad Ehsan, Soleimanpour-Lichaei Hamid Reza
Department of Stem Cells and Regenerative Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Gastroenterol Hepatol Bed Bench. 2023;16(1):492-498. doi: 10.22037/ghfbb.v16i1.2661.
This study aims to confirm previous fundamental in vitro findings about the PIWIL2 gene by investigating the effects of its overexpression on cell cycle, proliferation, apoptosis, and stem cell expression markers in colorectal cancer cells (CRC cells) at in vivo level.
PIWIL2 has a critical role in maintaining cellular stemness and proliferation. PIWIL2 is an oncogene whose expression in CRC is associated with the occurrence, metastasis, and poor prognosis.
SW480 cells harboring expression vectors with/without PIWIL2 were cultured and inoculated in BALB/c nude mice. Tumor formation and growth were monitored every 3 days. On the 28th day after inoculation, the tumors were harvested for their total RNA extraction, and the expression profiling of the candidate genes was performed by Real-time PCR.
Our results for the expression profiling of the xenografted tumors showed a significant increase in the expression of cancer stem cell markers, including CD24, CD133, and pluripotency marker SOX2 in the PIWIL2 over-expressing xenografts, compared to the control cell line. Moreover, PIWIL2 dramatically promoted the anti-apoptotic pathway by inducing STAT3 and BCL2-L1 genes in the PIWIL2 over-expressing xenografts, along with the up-regulation of Cyclin D1 and Ki-67 genes.
This research supports our prior in vitro findings, highlighting the critical role that PIWIL2 plays in the development of CRC and its substantial promise as a leading candidate for CRC-targeted therapy.
本研究旨在通过在体内水平研究PIWIL2基因过表达对结肠癌细胞（CRC细胞）的细胞周期、增殖、凋亡和干细胞表达标志物的影响，来证实先前关于该基因的基础体外研究结果。
PIWIL2在维持细胞干性和增殖中起关键作用。PIWIL2是一种癌基因，其在CRC中的表达与发生、转移及不良预后相关。
培养携带含/不含PIWIL2表达载体的SW480细胞，并接种于BALB/c裸鼠。每3天监测肿瘤形成和生长情况。接种后第28天，收获肿瘤用于提取总RNA，并通过实时PCR进行候选基因的表达谱分析。
我们对异种移植肿瘤的表达谱分析结果显示，与对照细胞系相比，PIWIL2过表达的异种移植瘤中癌症干细胞标志物（包括CD24、CD133和多能性标志物SOX2）的表达显著增加。此外，PIWIL2通过在PIWIL2过表达的异种移植瘤中诱导STAT3和BCL2-L1基因，以及上调细胞周期蛋白D1和Ki-67基因，显著促进抗凋亡途径。
本研究支持我们先前的体外研究结果，突出了PIWIL2在CRC发展中所起的关键作用及其作为CRC靶向治疗主要候选者的巨大潜力。
