Yang Hong, Pu Leilei, Li Ruobing, Zhu Rong
Department of Gastroenterology, Digestive Disease Hospital, Affiliated Hospital of Zunyi Medical University, Zunyi, China.
J Gastrointest Oncol. 2023 Apr 29;14(2):900-912. doi: 10.21037/jgo-23-63. Epub 2023 Feb 27.
Colorectal cancer (CRC) is highly heterogeneous at the genetic and molecular level and a major contributor to cancer-death worldwide. Non-structural maintenance of chromosomes (SMC) condensin I complex subunit G () is a subunit of condensin I and has been shown to be associated with the prognosis of cancers. This study investigated the functional role of in CRC and its mechanism.
Messenger RNA (mRNA) and protein expressions of and chromobox protein homolog 3 () were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot. The proliferation, cycle, and apoptosis of HCT116 cells were analyzed by Cell Counting Kit-8 (CCK-8), flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. RT-qPCR and western blot were used to determine the transfection efficacy of short hairpin (sh)-NCAPG and sh-CBX3. Western blot was used to explore cycle-, apoptosis-, and Wnt/β-catenin signaling-related proteins, and the activity of promoter was evaluated using a luciferase report assay. The expressions of cleaved caspase9 and cleaved caspase3 were assessed by colorimetric caspase activity assay.
The results showed that expression was elevated in CRC cells. After transfection with sh-NCAPG, expression was reduced. It was also discovered that knockdown suppressed proliferation and the cell cycle but induced apoptosis in HCT116 cells. The Human Transcription Factor Database (HumanTFDB; http://bioinfo.life.hust.edu.cn/HumanTFDB#!/) predicted the binding sites of and promoters. Meanwhile, the Encyclopedia of RNA Interactomes (ENCORI) database (https://starbase.sysu.edu.cn/) revealed that was positively correlated with . Our results showed that was transcriptionally regulated by . Additionally, Wnt/β-catenin signaling was discovered to be activated by overexpression. Further experiments showed that transcriptionally regulated by activated Wnt/β-catenin signaling to regulate the proliferation, cell cycle, and apoptosis of HCT116 cells.
Collectively, the results of our study indicated that was transcriptionally regulated by and activated the Wnt/β-catenin signaling pathway to facilitate the progression of CRC.
结直肠癌（CRC）在基因和分子水平上具有高度异质性，是全球癌症死亡的主要原因。染色体非结构维持（SMC）凝聚素I复合物亚基G（）是凝聚素I的一个亚基，已被证明与癌症预后相关。本研究探讨了在结直肠癌中的功能作用及其机制。
通过逆转录定量聚合酶链反应（RT-qPCR）和蛋白质印迹法检测和染色质盒蛋白同源物3（）的信使核糖核酸（mRNA）和蛋白质表达。采用细胞计数试剂盒-8（CCK-8）、流式细胞术和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）试验分析HCT116细胞的增殖、周期和凋亡。RT-qPCR和蛋白质印迹法用于测定短发夹（sh）-NCAPG和sh-CBX3的转染效率。蛋白质印迹法用于探索周期、凋亡和Wnt/β-连环蛋白信号相关蛋白，并使用荧光素酶报告试验评估启动子的活性。通过比色法半胱天冬酶活性试验评估裂解的半胱天冬酶9和裂解的半胱天冬酶3的表达。
结果显示，在结直肠癌细胞中表达升高。用sh-NCAPG转染后，表达降低。还发现敲低抑制HCT116细胞的增殖和细胞周期，但诱导其凋亡。人类转录因子数据库（HumanTFDB；http://bioinfo.life.hust.edu.cn/HumanTFDB#!/）预测了和启动子的结合位点。同时，RNA相互作用组百科全书（ENCORI）数据库（https://starbase.sysu.edu.cn/）显示与呈正相关。我们的结果表明受转录调控。此外，发现过表达激活Wnt/β-连环蛋白信号。进一步实验表明，受转录调控激活Wnt/β-连环蛋白信号，从而调节HCT116细胞的增殖、细胞周期和凋亡。
总的来说，我们的研究结果表明受转录调控，并激活Wnt/β-连环蛋白信号通路以促进结直肠癌的进展。
