Effect of DIAPH3 on proliferation, migration and invasion of gastric cancer cells and its molecular mechanism

贺国洋; 陈庆庆; 邓美静; 王高翔; 王贝玺; 王永霞; 李　巍; 千新来; 朱会芳

doi:10.19401/j.cnki.1007-3639.2021.12.005

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Effect of DIAPH3 on proliferation, migration and invasion of gastric cancer cells and its molecular mechanism

更新时间：2025-12-31

- Effect of DIAPH3 on proliferation, migration and invasion of gastric cancer cells and its molecular mechanism
- China Oncology Vol. 31, Issue 12, Pages: 1174-1184(2021)
- 作者机构：
  
  1. 新乡医学院病理学系,河南,新乡,453003
  2. 新乡医学院第三附属医院病理科,河南,新乡,453003
  3. 新乡医学院第一附属医院结直肠肛门外科,河南,新乡,453003
  4. 新乡医学院第四临床学院病理科,河南,新乡,453003
  5. 新乡医学院法医物证教研室,河南,新乡,453003
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2021.12.005
  CLC：
- Published Online：07 January 2022，
  
  Published：07 January 2021
- 稿件说明：
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贺国洋, 陈庆庆, 邓美静, 王高翔, 王贝玺, 王永霞, 李　巍, 千新来, 朱会芳, . DIAPH3对胃癌细胞增殖、迁移和侵袭的影响及分子机制[J]. 中国癌症杂志, 2021, 31(12): 1174-1184.

贺国洋, 陈庆庆, 邓美静, et al. Effect of DIAPH3 on proliferation, migration and invasion of gastric cancer cells and its molecular mechanism[J]. China Oncology, 2021, 31(12): 1174-1184.
贺国洋, 陈庆庆, 邓美静, 王高翔, 王贝玺, 王永霞, 李　巍, 千新来, 朱会芳, . DIAPH3对胃癌细胞增殖、迁移和侵袭的影响及分子机制[J]. 中国癌症杂志, 2021, 31(12): 1174-1184. DOI： 10.19401/j.cnki.1007-3639.2021.12.005.

贺国洋, 陈庆庆, 邓美静, et al. Effect of DIAPH3 on proliferation, migration and invasion of gastric cancer cells and its molecular mechanism[J]. China Oncology, 2021, 31(12): 1174-1184. DOI： 10.19401/j.cnki.1007-3639.2021.12.005.

摘要

背景与目的：胃癌是消化系统常见的恶性肿瘤之一，但其发病机制尚不清楚。Diaphanous相关成蛋白3（diaphanous-related formin 3，DIAPH3）对多种肿瘤的发生、发展具有重要作用，但其在胃癌中的作用未见报道。探讨DIAPH3在胃癌组织中的表达及其对胃癌细胞增殖、迁移和侵袭的影响及其分子机制。方法：利用基因表达谱分析（gene expression profiling interactive analysis，GEPIA）数据库在线分析DIAPH3在胃癌组织中的表达；收集2020年1月—2020年12月年新乡医学院第四临床学院病理科保存的62例胃癌患者的石蜡包埋组织及配对癌旁组织标本，采用免疫组织化学染色方法检测胃癌组织中DIAPH3的表达，并进行临床病理学相关性分析；采用蛋白质印迹法（Western blot）检测敲低或过表达DIAPH3后对DIAPH3、细胞周期蛋白D1（cyclin D1）、E-钙黏蛋白（E-cadherin）、波形蛋白（vimentin）及N-钙黏蛋白（N-cadherin）蛋白质水平的变化。采用实时荧光定量聚合酶链反应（real-time fluorescent quantitative polymerase chain reaction，RTFQ-PCR）检测敲低或过表达DIAPH3后对DIAPH3转录水平表达的影响。利用细胞计数试剂盒-8（cellcounting kit-8，CCK-8）实验检测细胞增殖；利用划痕愈合实验检测细胞迁移；利用transwell小室实验检测细胞侵袭。结果：GEPIA数据库在线分析显示，与胃癌癌旁组织相比，DIAPH3 mRNA在胃癌组织中表达升高（P ＜ 0.05）；胃癌组织中DIAPH3的阳性表达率为70.97%（44/62），高于胃癌癌旁组织16.13%（10/62），差异有统计学意义（P ＜ 0.01）；与高中分化组比较，低分化组DIAPH3表达升高（P ＜ 0.05）；与无淋巴结转移组相比，有淋巴结转移组DIAPH3表达升高（P ＜ 0.05）。干扰DIAPH3组细胞增殖活力、细胞迁移率和细胞侵袭数均低于阴性对照组（P ＜ 0.05）；过表达DIAPH3组细胞增殖活力、细胞迁移率和细胞侵袭数均高于过表达对照组（P ＜ 0.05）。过表达DIAPH3后干扰cyclin D1，胃癌细胞株的增殖活力低于过表达DIAPH3组，高于过表达对照组（P ＜ 0.05）。过表达DIAPH3后干扰vimentin，胃癌细胞株的细胞迁移率和细胞侵袭数低于过表达DIAPH3组，高于过表达对照组（P ＜ 0.05）。与干扰对照组相比，干扰DIAPH3组E-cadherin蛋白质表达增加，DIAPH3、cyclin D1、vimentin和N-cadherin蛋白质表达降低（P ＜ 0.05）；与过表达对照组相比，过表达DIAPH3组中E-cadherin蛋白质表达降低，DIAPH3、cyclin D1、vimentin和N-cadherin蛋白质表达增加（P ＜ 0.05）。在敲低或过表达DIAPH3胃癌细胞株中，DIAPH3在mRNA水平均显著降低或升高（P ＜ 0.05）。结论：DIAPH3可促进胃癌细胞的增殖、迁移和侵袭，其作用与上调cyclin D1、促进上皮-间充质转化有关。

Abstract

Background and purpose: Gastric cancer is one of the common malignant tumors in the digestive system

but its pathogenesis is not clear. Diaphanous-related formin 3 (DIAPH3) plays an important role in the occurrence and development of a variety of tumors

however

its role in gastric cancer has not been reported. This study was to investigate the expression of DIAPH3 in gastric cancer and its effect on the proliferation

migration and invasion of gastric cancer cells. Methods: Gene expression profiling interactive analysis (GEPIA) database was used to analyze the expression of DIAPH3 in gastric cancer. Paraffin embedded tissues and paired adjacent tissues from 62 patients with gastric cancer were collected. The expression of DIAPH3 in gastric cancer was detected by immunohistochemistry

and the clinicopathological correlation was analyzed. Western blot was performed to detect the effects of knockdown or over-expression of DIAPH3 on the protein levels of DIAPH3

cyclin D1

E-cadherin

vimentin and N-cadherin. Real- time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the effect of knockdown or over-expression of DIAPH3 mRNA. Cell proliferation was detected by cell counting kit-8 (CCK-8). Cell migration was detected by the wound healing assay

and cell invasion was detected by transwell chamber experiment. Results: GEPIA database online predicted that the expression of DIAPH3 mRNA was higher in gastric cancer than in adjacent non-cancer tissues (P＜0.05). The positive expression rate of DIAPH3 in gastric cancer was 70.97% (44/62)

which was higher than that in adjacent non-cancer tissues (16.13%

10/62

P＜0.01). Compared with the high differentiation group

the expression of DIAPH3 in the low differentiation group was higher (P＜0.05). Compared with the group without lymph node metastasis

the expression of DIAPH3 in the group with lymph node metastasis was higher (P＜0.05). The cell proliferation activity

cell migration rate and cell invasion were lower in the interference DIAPH3 group than in the negative control group (P＜0.05). The cell proliferation activity

cell migration rate and cell invasion number were higher in the over-expression of DIAPH3 group than in the over-expression control group (P＜0.05). Following over- expression of DIAPH3 and interference of cyclin D1

the proliferation activity of gastric cancer cell line was lower compared with over-expression of DIAPH3 group and higher compared with over-expression control group (P＜0.05). Following over-expression of DIAPH3 and interference of vimentin

the cell migration rate and cell invasion number of gastric cancer cell line were lower compared with over-expression DIAPH3 group and higher compared with over-expression control group (P＜0.05). Compared with the interference control group

the expression of E-cadherin protein increased

and the expressions of DIAPH3

cyclin D1

vimentin and N-cadherin decreased in the interference DIAPH3 group (P＜0.05). Compared with the control group

the expression of E-cadherin protein decreased

and the protein expressions of DIAPH3

cyclin D1

vimentin and N-cadherin increased in the over-expression group. In knockdown or over-expression of DIAPH3 gastric cancer cell lines

the mRNA level of DIAPH3 decreased or increased significantly (P＜0.05). Conclusion: DIAPH3 promotes the proliferation

migration and invasion of gastric cancer cells. The role of DIAPH3 is associated with up-regulation of cyclin D1 and epithelial-mesenchymal transition.

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