Bcl-2转录抑制因子1在胃癌中的表达及其生物学功能
作者:
作者单位:

1.湖北医药学院 药学院,湖北 十堰 442000;2.湖北医药学院附属太和医院 肝胆胰外科,湖北 十堰 442000

作者简介:

赵馨旭,湖北医药学院药学院/湖北医药学院附属太和医院硕士研究生,主要从事药理学、分子生物学方面的研究。

通信作者:

李飞,Email: piaopodexinlifei@163.com

基金项目:

湖北省卫健委青年人才基金资助项目(WJ2021Q010);湖北省教育厅科学研究计划基金资助项目(D20192101);湖北省十堰市科技局引导性科研基金资助项目(19Y13);湖北医药学院人才启动金基金资助项目(2018QDJZR29)。


Expression of Bcl-2 transcription inhibitor 1 in gastric cancer and its biological function
Author:
Affiliation:

1.School of Pharmaceutic Science, Hubei University of Medicine, Shiyan, Hubei 442000, China;2.Department of Hepatopancreatobiliary Surgery, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 音频文件
  • |
  • 视频文件
    摘要:

    背景与目的 失巢凋亡效应分子Bcl-2转录抑制因子1(Bit1)在胃癌组织中存在异常表达,Bit1在胃癌中功能及作用机制如何值得研究。本研究旨在进一步观察Bit1在胃癌组织中的表达,并初步分析其对胃癌细胞生物学行为的影响。方法 用Western blot检测20例原发性胃癌患者癌组织及癌旁组织手术标本中Bit1蛋白的表达;将胃癌BGC-803细胞分别转染Bit1 shRNA慢病毒载体或空病毒载体后,以无处理的野生型BGC-803细胞为空白对照,用细胞免疫荧光实验验证转染效率与Bit1沉默效果,并分别用细胞划痕实验、Transwell侵袭实验、细胞增殖实验及TUNEL实验检测细胞侵袭、迁移、增殖及凋亡情况。结果 Western blot结果显示,与对应癌旁组织比较,Bit1在胃癌组织中蛋白表达水平明显上调(P<0.01)。免疫荧光结果显示,转染成功且Bit1被有效沉默。细胞行为学实验显示,与空白对照细胞比较,转染Bit1 shRNA后的BGC-803细胞的划痕愈合率明显降低、侵袭细胞数量明显减少、细胞增殖活性明显减弱、晚期凋亡细胞数明显增加(均P<0.05);转染空载体的BGC-803细胞以上细胞行为学指标与空白对照细胞间的差异均无统计学意义(均P>0.05)。结论 Bit1在胃癌中表达上调,并与胃癌细胞的恶性生物学行为密切相关,其有可能是与胃癌发生、发展、预后相关联的生物标志物及潜在治疗靶点。

    Abstract:

    Background and Aims Anoikis effector Bcl-2 transcription inhibitor 1 (Bit1) is abnormally expressed in gastric cancer tissue, the function and mechanism of Bit1 in gastric cancer is worth studying. This study was conducted to further examine the expression of Bit1 in gastric cancer tissue, and present a preliminary analysis of the effects of Bit1 on the biological behavior of gastric cancer cells.Methods The expressions of Bit1 in the surgical specimens of gastric cancer and corresponding adjacent tissue from 20 patients with primary gastric cancer were determined by Western blot analysis. In gastric cancer BGC-803 cells after transfection with Bit1 shRNA lentivirus or empty lentivirus vector, with the untreated wild type BGC-803 cells as blank control, the transfection efficiency and Bit1 silencing effect were validated by immunofluorescence assay, and then, the invasion, migration, proliferation and apoptosis were Transwell invasion assay, scratch wound assay, CCK-8 assay and TUNEL staining assay, respectively.Results The results of Western blot showed that the expression level of Bit1 protein in gastric cancer tissue was significantly up-regulated compared with corresponding adjacent tissues (P<0.01). The results of immunofluorescence assay demonstrated the successful transfection and effective Bit1 silencing. The results of series of cellular behavior experiments revealed that the wound-healing rate, number of invaded cells, and proliferation ability were all significantly decreased, while the number of late apoptotic cells was significantly increased in BGC-803 cells with Bit1 shRNA transfection compared with blank control cells (all P<0.05); there were no statistical differences in these cellular behavior parameters between BGC-803 cells transfected with empty vector and blank control cells (all P>0.05).Conclusion The expression level of Bit1 is up-regulated in gastric cancer, and it is closely related to the malignant biological behavior of gastric cancer cells, suggesting that Bit1 may be a biomarker associated with the initiation, progression and prognosis as well as a candidate therapeutic target for gastric cancer.

    图1 Western blot检测Bit1蛋白表达 A:20例胃癌患者癌组织与癌旁组织Bit1蛋白表达;B-C:Bit1蛋白在癌组织与癌旁组织中的相对表达量Fig.1 Expressions of Bit1 protein detected by Western blot A: Expressions of Bit1 protein in gastric cancer and corresponding adjacent tissues from 20 gastric cancer patients; B-C: Relative expression levels of Bit1 protein in gastric cancer and corresponding adjacent tissues
    图2 免疫荧光检测 A:转染效率检测(×100);B:Bit1沉默效果检测(×1 000)Fig.2 Immunofluorescence staining assay A: Transfection efficiency determination (×100); B: Silencing effect detection (×1 000)
    图3 划痕实验检测各组BGC-803细胞的迁移能力Fig.3 Migration ability of each group of BGC-803 cells measured by scratch wound assay
    图4 Transwell实验检测各组BGC-803细胞的侵袭能力Fig.4 Invasion ability of each group of BGC-803 cells determined by Transwell assay
    图5 CCK-8法检测各组BGC-803细胞的增殖活力Fig.5 Proliferation ability of each group of BGC-803 cells analyzed by CCK-8 assay
    图6 TUNEL法检测各组BGC-803细胞的凋亡情况Fig.6 Apoptosis in each group of BGC-803 cells examined by TUNEL staining assay
    参考文献
    相似文献
    引证文献
引用本文

赵馨旭,黄林生,刘佳玲,韩俊祥,余惠凡,李飞. Bcl-2转录抑制因子1在胃癌中的表达及其生物学功能[J].中国普通外科杂志,2022,31(4):481-489.
DOI:10.7659/j. issn.1005-6947.2022.04.010

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
历史
  • 收稿日期:2021-09-22
  • 最后修改日期:2021-12-02
  • 录用日期:
  • 在线发布日期: 2022-05-07