Luo H, Pan Y, Chen W, Zhang W, Shao S, Yang Q, Li T
College of Life Science and Agroforestry, Qiqihar University, Qiqihar 161006, China.
Key Laboratory of Resistant Genetic Engineering and Cold Biodiversity Conservation of Heilongjiang Province, Qiqihar 161006, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Aug 20;42(8):1126-1133. doi: 10.12122/j.issn.1673-4254.2022.08.03.
To investigate the effect of interference of CTPS gene on toosendanin-induced apoptosis of gastric cancer MKN-45 cells.
Bioinformatic analysis was used to analyze CTPS gene expression in human gastric cancer tissues and the overall survival of gastric cancer patients with high CTPS gene expression. Human gastric cancer MKN-45 cells were transfected with a short hairpin interfering RNA targeting CTPS gene, and 48 h later, qRT-PCR and Western blotting were used to detect cellular expression CTPS at both the mRNA and protein levels. MKN-45 cells with CTPS knockdown were treated with 80 nmol/L toosendanin for 48 h, and the cell viability was assessed with MTT assay; the cell morphology was observed using laser confocal microscope, and the expression of γH2AX was detected with immunofluorescence assay.
Bioinformatic analysis suggested that CTPS was highly expressed in human gastric cancer tissues, and gastric cancer patients with high CTPS gene expression had a shorter overall survival. MKN-45 cells transfected with Sh-CTPS interference vector showed significantly lowered cell survival rate ( < 0.01) with obvious cell shrinkage, irregular morphology, typical apoptotic changes, and increased cell apoptosis rate ( < 0.05). Treatment of the transfected cells with 80 nmol/L toosendanin for 48 h resulted in further reduction of the cell survival rate ( < 0.001), and the cells showed an increased apoptotic rate ( < 0.05) with appearance of apoptotic bodies.
Interference of CTPS gene can promote TSN-induced apoptosis of gastric cancer MKN-45 cells.
探讨CTPS基因干扰对川楝素诱导胃癌MKN-45细胞凋亡的影响。
采用生物信息学分析方法分析CTPS基因在人胃癌组织中的表达情况以及CTPS基因高表达的胃癌患者的总生存期。将靶向CTPS基因的短发夹干扰RNA转染人胃癌MKN-45细胞，48 h后，采用qRT-PCR和蛋白质印迹法分别从mRNA和蛋白质水平检测细胞中CTPS的表达。用80 nmol/L川楝素处理CTPS基因敲低的MKN-45细胞48 h，采用MTT法评估细胞活力；用激光共聚焦显微镜观察细胞形态，采用免疫荧光法检测γH2AX的表达。
生物信息学分析提示CTPS在人胃癌组织中高表达，CTPS基因高表达的胃癌患者总生存期较短。转染Sh-CTPS干扰载体的MKN-45细胞存活率显著降低（<0.01），细胞明显皱缩，形态不规则，出现典型的凋亡改变，细胞凋亡率升高（<0.05）。用80 nmol/L川楝素处理转染后的细胞48 h，细胞存活率进一步降低（<0.001），细胞凋亡率升高（<0.05），出现凋亡小体。
CTPS基因干扰可促进川楝素诱导的胃癌MKN-45细胞凋亡。
