Department of Gastroenterology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, No. 107 Yanjiang West Road, Guangzhou, 510120, Guangdong, People's Republic of China.
Dig Dis Sci. 2013 Sep;58(9):2518-27. doi: 10.1007/s10620-013-2752-4. Epub 2013 Jul 6.
BACKGROUND/AIMS: Pancreatic cancer has the worst prognosis of any gastrointestinal cancer with a mortality rate approaching its incidence. Previous studies have indicated that GATA6 plays a key role in organ development and function, and that abnormal expression of GATA6 may induce tumorigenesis. Meanwhile, it has been reported that generation of reactive oxygen species contributes to carcinogenesis. In this study, we set out to study the role of GATA6 expression on proliferation and apoptosis of pancreatic cancer cells and the role of reactive oxygen species.
Four target miRNA sequences against GATA6 mRNA were synthesized and used to transfect SW1990 cells. Then, GATA6 expression in SW1990 cells was examined by western blot and quantative real-time polymerase chain reaction. Cell proliferation was examined by WST-8 and colony formation assay. Cell cycle progression and apoptosis were measured by flow cytometry. We also measured the generation of reactive oxygen species by immunofluorescence and flow cytometry.
RNA interference against GATA6 successfully inhibited mRNA and protein expression of GATA6 in the SW1990 pancreatic cancer cell line. Silencing of GATA6 by RNA interference inhibited cell proliferation and increased apoptosis of SW1990, and enhanced the expression of reactive oxygen species.
These results suggest that the RNA interference approach against GATA6 may be an effective therapeutic approach for treatment of pancreatic cancer.
背景/目的：胰腺癌的预后是所有胃肠道癌中最差的，其死亡率接近发病率。先前的研究表明，GATA6 在器官发育和功能中发挥着关键作用，GATA6 的异常表达可能诱导肿瘤发生。同时，有报道称活性氧的产生有助于致癌作用。在本研究中，我们旨在研究 GATA6 表达对胰腺癌细胞增殖和凋亡的作用以及活性氧的作用。
合成了针对 GATA6 mRNA 的四个靶 miRNA 序列，并用于转染 SW1990 细胞。然后，通过 Western blot 和定量实时聚合酶链反应检测 SW1990 细胞中 GATA6 的表达。通过 WST-8 和集落形成测定法检测细胞增殖。通过流式细胞术测量细胞周期进程和细胞凋亡。我们还通过免疫荧光和流式细胞术测量活性氧的产生。
针对 GATA6 的 RNA 干扰成功抑制了 SW1990 胰腺癌细胞系中 GATA6 的 mRNA 和蛋白表达。RNA 干扰沉默 GATA6 抑制了 SW1990 的细胞增殖并增加了细胞凋亡，并增强了活性氧的表达。
这些结果表明，针对 GATA6 的 RNA 干扰方法可能是治疗胰腺癌的有效治疗方法。
