Chen Zhixiong, Liu Banglun, Zhang Shouru, Chen Lihui, Lv Yuyu, Sun Hao
Department of Gastrointestinal Cancer Center, Chongqing University Cancer Hospital, Chongqing, China.
Department of Pathology, Chongqing University Cancer Hospital, Chongqing, China.
Turk J Biol. 2022 Apr 25;46(4):277-287. doi: 10.55730/1300-0152.2616. eCollection 2022.
Gastric cancer is becoming the 4th leading cause of cancer-associated death worldwide. The purpose of this study was to investigate the role of RGS1 in gastric cancer in vitro and in vivo. Proliferation, migration, invasion, and colony formation of NCIN87 cells and drug-resistant NCIN87 cells (NCIN87-DR) were determined. Cell apoptosis and cell cycle were examined using a flow cytometry assay. RGS1 gene knock-down vector (pLVshshRGS1) and Xenograft tumor mouse model was generated. RGS1 and epithelial-mesenchymal transition (EMT) associated markers, including E-cadherin (E-cad), N-cadherin (N-cad), Slug, and Vimentin were detected using a western blotting assay. Tumor size of Xenograft tumor mouse was measured and Ki67 expression was detected using the immunohistochemical assay. NCIN87-DR cells demonstrated significantly lower proliferation, migration, and invasion compared to those of NCIN87 cells (p < 0.05). NCIN87-DR cells showed obvious early apoptosis and displayed obvious alterations for the cell cycle. NCIN87-DR cells exhibited predominantly higher RGS1 expression than that in NCIN87 cells (p < 0.01). E-cad expression was markedly decreased (p < 0.01) and N-cad (p < 0.05), Slug (p < 0.01), Vimentin (p < 0.05) expressions were significantly increased in NCIN87-DR cells than those in NCIN87 cells. RGS1 gene silence remarkably reduced NCIN87-DR proliferation compared to that in NCIN87-DR cells without treatment (p < 0.01). RGS1 gene-silenced NCIN87-DR cell immunization predominantly inhibited tumor growth in Xenograft tumor mouse than that without RGS1 silence (p < 0.05). RGS1 gene-silenced NCIN87-DR cell immunization significantly downregulated Ki67 expression in tumor tissues compared with that without RGS1 silence. In conclusion, RGS1 gene silence reduced the proliferation of NCIN87-DR cells in vitro and inhibited tumor growth in vivo. Therefore, RGS1 served as an antitumor target for the gastric cancer treatment.
胃癌正成为全球癌症相关死亡的第四大主要原因。本研究的目的是在体外和体内研究RGS1在胃癌中的作用。测定了NCIN87细胞和耐药NCIN87细胞（NCIN87-DR）的增殖、迁移、侵袭和集落形成能力。使用流式细胞术检测细胞凋亡和细胞周期。构建了RGS1基因敲低载体（pLVshshRGS1）和异种移植肿瘤小鼠模型。使用蛋白质免疫印迹法检测RGS1和上皮-间质转化（EMT）相关标志物，包括E-钙黏蛋白（E-cad）、N-钙黏蛋白（N-cad）、锌指蛋白Slug和波形蛋白。测量异种移植肿瘤小鼠的肿瘤大小，并使用免疫组织化学法检测Ki67表达。与NCIN87细胞相比，NCIN87-DR细胞的增殖、迁移和侵袭能力显著降低（p<0.05）。NCIN87-DR细胞表现出明显的早期凋亡，并且细胞周期出现明显改变。NCIN87-DR细胞中RGS1的表达明显高于NCIN87细胞（p<0.01）。与NCIN87细胞相比，NCIN87-DR细胞中E-cad的表达明显降低（p<0.01），而N-cad（p<0.05）、Slug（p<0.01）、波形蛋白（p<0.05）的表达显著增加。与未处理的NCIN87-DR细胞相比，RGS1基因沉默显著降低了NCIN87-DR细胞的增殖（p<0.01）。与未沉默RGS1的情况相比，RGS1基因沉默的NCIN87-DR细胞免疫接种主要抑制了异种移植肿瘤小鼠的肿瘤生长（p<0.05）。与未沉默RGS1的情况相比，RGS1基因沉默的NCIN87-DR细胞免疫接种显著下调了肿瘤组织中Ki67的表达。总之，RGS1基因沉默降低了NCIN87-DR细胞在体外的增殖，并抑制了体内肿瘤生长。因此，RGS1可作为胃癌治疗的抗肿瘤靶点。
