The influence of silencing<em>leptin</em>gene on the invasion and migration abilities of gallbladder cancer GBC-SD and OCUG cells

石万红; 邹　雷; 康　强; 王峻峰; 白建华; 晋　云; 张　杰; 张小文

doi:10.19401/j.cnki.1007-3639.2021.11.002

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The influence of silencingleptingene on the invasion and migration abilities of gallbladder cancer GBC-SD and OCUG cells

更新时间：2025-12-31

- The influence of silencingleptingene on the invasion and migration abilities of gallbladder cancer GBC-SD and OCUG cells
- China Oncology Vol. 31, Issue 11, Pages: 1050-1057(2021)
- 作者机构：
  
  1. 昆明医科大学第一附属医院器官移植科,云南,昆明,650032
  2. 昆明医科大学第二附属医院肝胆外科,云南,昆明,650101
  3. 云南省第一人民医院肝胆外科,云南,昆明,650021
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2021.11.002
  CLC：
- Published Online：02 December 2021，
  
  Published：02 December 2021
- 稿件说明：
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石万红, 邹　雷, 康　强, 王峻峰, 白建华, 晋　云, 张　杰, 张小文 .

石万红, 邹　雷, 康　强, et al. The influence of silencingleptingene on the invasion and migration abilities of gallbladder cancer GBC-SD and OCUG cells[J]. China Oncology, 2021, 31(11): 1050-1057.
石万红, 邹　雷, 康　强, 王峻峰, 白建华, 晋　云, 张　杰, 张小文 . DOI： 10.19401/j.cnki.1007-3639.2021.11.002.

石万红, 邹　雷, 康　强, et al. The influence of silencingleptingene on the invasion and migration abilities of gallbladder cancer GBC-SD and OCUG cells[J]. China Oncology, 2021, 31(11): 1050-1057. DOI： 10.19401/j.cnki.1007-3639.2021.11.002.

摘要

背景与目的：研究表明，与肥胖症密切相关的源自脂肪细胞的细胞因子瘦素（leptin）在癌变和肿瘤发生中起着重要作用。通过体外实验观察leptin对人胆囊癌细胞系GBC-SD和OCUG侵袭和迁移能力的影响，探讨其可能的作用机制。方法：将胆囊癌GBC-SD和OCUG细胞分为对照组和实验组。实验组利用小干扰RNA（small interfering RNA，siRNA）技术靶向沉默胆囊癌GBC-SD和OCUG细胞的leptin表达，应用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测转染效率，通过细胞划痕实验和transwell小室法检测胆囊癌GBC-SD和OCUG细胞的迁移和侵袭能力，采用免疫荧光和免疫细胞化学实验检测siRNA干扰后胆囊癌GBC-SD和OCUG细胞的leptin蛋白水平，利用蛋白质印迹法（Western blot）检测干扰后GBC-SD和OCUG细胞的leptin和p-AKT蛋白表达。结果：细胞划痕实验结果显示，利用siRNA靶向沉默leptin后GBC-SD和OCUG细胞的迁移能力下降（t=26.614，P ＜ 0.01；t=19.338，P ＜ 0.01）。Transwell迁移实验结果显示，利用siRNA靶向沉默leptin后GBC-SD和OCUG细胞的迁移能力也下降（t=7.185，P=0.002；t=8.889，P=0.003）。Transwell侵袭实验结果显示，干扰后GBC-SD和OCUG细胞的侵袭能力下降（t=10.183，P=0.001；t=9.697，P=0.001）。免疫荧光和免疫细胞化学实验结果显示，干扰后GBC-SD和OCUG细胞的leptin蛋白表达下降，Westernblot检测结果表明，靶向沉默GBC-SD细胞后leptin和p-AKT蛋白表达下调（t=26.463，P ＜ 0.01；t=13.904，P ＜ 0.01），靶向沉默OCUG细胞后leptin和p-AKT蛋白表达也下调（t=21.335，P ＜ 0.01；t=17.914，P ＜ 0.01）。结论：下调GBC-SD和OCUG细胞的leptin表达可抑制细胞侵袭和转移能力，并可能通过AKT信号通路调控胆囊癌细胞的侵袭和迁移。Leptin有望成为胆囊癌治疗的一个重要靶点。

Abstract

Background and purpose: Emerging evidence suggests that fat cell-derived cytokine leptin

which is closely related to obesity

plays an important role in carcinogenesis and tumorigenesis. In this study

the effect of leptin on the invasion and migration abilities of GBC-SD and OCUG cells were observed in vitro

and its possible related mechanisms were explored. Methods: GBC-SD and OCUG cells were divided into control group and experimental group. In the experimental group

we used small interfering RNA (siRNA) to target and silence the expression of leptin in GBC-SD and OCUG cells. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the transfection efficiency. Wound healing assay and transwell assay were used to detect the migration and invasion abilities of GBC-SD and OCUG cells. The immunofluorescence and immunocytochemistry experiments were used to detect the expression of leptin protein in GBC-SD and OCUG cells after siRNA interference. The protein expressions of leptin and p-AKT in GBC-SD and OCUG cells after interference were detected by Western blot. Results: The results of wound healing assay showed that the migration abilities of GBC-SD and OCUG cells decreased after silencing leptin with siRNA interference (t=26.614

P ＜ 0.01; t=19.338

P ＜ 0.01). The results of transwell migration assay also showed that the migration abilities of GBC-SD and OCUG cells decreased after silencing leptin with siRNA interference (t=7.185

P=0.002; t=8.889

P=0.003). Transwell invasion assay results showed that the invasion abilities of GBC-SD and OCUG cells decreased after interference (t=10.183

P=0.001; t=9.697

P=0.001). Immunofluorescence and immunocytochemistry experiments showed decreased leptin protein expression in GBC-SD and OCUG cells after interference. Western blot showed that leptin and p-AKT protein expressions were down-regulated after targeted silencing in GBC-SD cells (t=26.463

P ＜ 0.01; t=13.904

P ＜ 0.01). Western blot showed that leptin and p-AKT protein expressions were also down-regulated after targeted silencing in OCUG cells (t=21.335

P ＜ 0.01; t=17.914

P ＜ 0.01). Conclusion: Down-regulation of leptin expression in GBC-SD and OCUG cells can inhibit cell invasion and metastasis

and may regulate gallbladder cancer invasion and migration through AKT signaling pathway. Leptin are expected to become an important target for gallbladder cancer treatment.

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The influence of silencingleptingene on the invasion and migration abilities of gallbladder cancer GBC-SD and OCUG cells

DOI：10.19401/j.cnki.1007-3639.2021.11.002

摘要

Abstract

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