1.武汉大学中南医院腹部肿瘤放化疗科,武汉 430071;2.武汉大学泰康医学院(基础医学院)病理教研室,武汉 43007
R-33
1.Department of Medical Oncology and Radiotherapy, Abdominal Tumor Division, Zhongnan Hospital of Wuhan University, Wuhan 430071, China. 2. Wuhan University TaiKang Medical School (School of Basic Medical Sciences), Wuhan 430071
目的 探讨层粘连蛋白γ2(LAMC2)通过PI3K/ Akt 对胃癌细胞增殖的影响。 方法 利用癌症基因组图谱(TCGA)及基因表达综合数据库(GEO)联合分析胃癌中与细胞增殖相关差异基因,根据差异倍数确定候选基因LAMC2=。UALCAN 及Kaplan-Meier plotter 在线数据库分析LAMC2 在胃癌中的表达及其与临床病理特征的关系,采用蛋白印迹(Western blot)法检测人胃癌细胞株(HGC-27、NCI-N87、SNU-1)中LAMC2 的表达。构建LAMC2敲减胃癌细胞模型,Western blot 检测LAMC2 的表达,确定模型构建效果。将HGC-27 及NCI-N87 细胞分成空白对照(Control)组、阴性对照(si-NC) 组及LAMC2 敲减(si-LAMC2) 组,5-乙炔基-2’ 脱氧尿嘧啶核苷(5-ethynyl-2’- deoxyuridine,EdU)及平板克隆实验分析细胞增殖及克隆形成能力,Western blot 检测细胞增殖蛋白(CyclinD1、PCNA、c-Myc)表达及PI3K/ Akt 通路激活情况。 结果 LAMC2 是胃癌中与细胞增殖相关的差异倍数最大的差异基因,其在胃癌中的表达显著上调,与胃癌的分期、分级及生存等临床病理相关。LAMC2 在人胃癌细胞(HGC-27、NCI-N87、SNU-1)中的表达均显著高于GES-1 细胞(均P<0. 01)。与si-NC 组比较,si-LAMC2 显著抑制HGC-27 及NCI-N87 细胞增殖,降低细胞克隆形成率(P<0. 01),细胞增殖蛋白(CyclinD1、PCNA 及c-Myc)表达均被显著降低(均P<0. 01),PI3Kp85 及Akt 磷酸化水平均被显著下调(均P<0. 01)。 结论 LAMC2 在胃癌中的表达上调,与胃癌的分期、分级及生存等临床病理相关,其可通过激活PI3K/ Akt 通路促进细胞增殖。
Objective To investigate the effect of laminin subunit γ2(LAMC2) on the proliferation of gastric cancer cells through the PI3K/ Akt pathway. Methods Differentially expressed genes related to gastric cancer cell proliferation were analyzed in the cancer genome atlas and gene expression omnibus databases. The candidate gene LAMC2 was identified in accordance with the multiple of differential. UALCAN and Kaplan-Meier Plotter online databases were used to analyze LAMC2 expression in gastric cancer and its relationship with clinicopathological features. Western blot was used to detect LAMC2 in human gastric cancer cell lines HGC-27, NCI-N87 and SNU-1. An LAMC2 knockout gastric cancer cell model was established, and LAMC2 was detected by Western blot to determine the effects of model establishment. HGC-27 and NCI-N87 cells were divided into blank control(Control), negative control group(si-NC), and LAMC2 knockout(si-LAMC2)group. 5-Ethynyl-2’-deoxyuridine and colony formation assays were used to evaluate cell proliferation and clonality. Western blot was used to assess expression of cell proliferation-related proteins(Cyclin D1, PCNA and c-Myc)and activation of the PI3K/ Akt pathway. Results LAMC2 was the most differentially expressed gene associated with cell proliferation in gastric cancer, and its expression was significantly upregulated in gastric cancer, which correlated to the clinicopathological characteristics of the gastric cancer stage, grade, and survival. The expression of LAMC2 in human gastric cancer cells (HGC-27, NCI-N87, SNU-1) was significantly higher than that in GES-1 cells(all P< 0. 01). Compared with the si-NC group, si-LAMC2 significantly inhibited HGC-27 and NCI-N87 cell proliferation, decreased the colony formation rate(P< 0. 01), and significantly decreased the expression of cell proliferation-related proteins(CyclinD1, PCNA and c-Myc) (all P< 0. 01). PI3Kp85 and Akt phosphorylation levels were also significantly downregulated( all P< 0. 01). Conclusions Upregulation of LAMC2 expression in gastric cancer is related to the clinicopathological characteristics of the gastric cancer stage, grade, and survival, and promotes cell proliferation by activation of the PI3K/ Akt pathway.
王文博,汪清云,郭卫,夏泠. LAMC2 通过PI3K/ Akt 信号通路调控胃癌细胞增殖的研究[J].中国比较医学杂志,2023,33(10):61~70.
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