β-防御素-1通过环状RNA HAS-CIRCpedia-5280抑制小鼠结肠癌的研究

doi:10.12160/j.issn.1672-5190.2026.03.001

摘要/Abstract

摘要： [目的] 借助裸鼠皮下成瘤体内实验,验证β-防御素-1通过环状RNA HAS-CIRCpedia-5280发挥抑制结肠癌肿瘤生长的作用。[方法] 采用慢病毒转导技术将携带HAS-CIRCpedia-5280的过表达慢病毒与空载体对照慢病毒分别转导人结肠癌HCT-116细胞,经聚凝胺辅助感染及嘌呤霉素连续筛选2周,构建HAS-CIRCpedia-5280过表达稳转HCT-116细胞株（HAS-CIRCpedia-5280细胞）与空载体对照稳转HCT-116细胞株（NC细胞）。将30只BALB/c-nu裸鼠分为6组[oe-NC组,皮下注射NC细胞;oe-circRNA组,皮下注射HAS-CIRCpedia-5280细胞;oe-NC+PBS组,皮下注射NC细胞,腹腔注射PBS;oe-circRNA+PBS组,皮下注射HAS-CIRCpedia-5280细胞,腹腔注射PBS;oe-NC+hBD-1（人β-防御素-1）组,皮下注射NC细胞,腹腔注射hBD-1;oe-circRNA+hBD-1组,皮下注射HAS-CIRCpedia-5280细胞,腹腔注射hBD-1],每组5只,皮下注射上述2种稳转细胞建立BALB/c-nu裸鼠皮下异种移植瘤模型,并对oe-NC+hBD-1组和oe-circRNA+hBD-1组、oe-NC+PBS组和oe-circRNA+PBS组分别腹腔注射0.1 mg/（kg·BW）的hBD-1或等体积的PBS予以干预,每周2次（周一和周四）,每7 d记录肿瘤体积。自首次给药起4周内完成全部注射,末次给药72 h后对裸鼠实施安乐死,并剥离肿瘤组织,测定肿瘤体积及重量。[结果] 通过慢病毒转导构建 HAS-CIRCpedia-5280 过表达（oe-circRNA）及空载体对照（oe-NC）稳转HCT-116细胞株,荧光显微镜观察可见,oe-circRNA 组和 oe-NC 组视野内多数细胞呈现清晰绿色荧光,细胞阳性率较高;qRT-PCR 结果显示,相较于 oe-NC 对照组,oe-circRNA 组 HAS-CIRCpedia-5280 表达水平极显著上调（P<0.01）。构建结肠癌裸鼠皮下移植瘤模型,定期监测肿瘤生长,结果显示,相较于 oe-NC 组,oe-circRNA组移植瘤重量极显著升高（P<0.01）、体积显著增大（P<0.05）;qRT-PCR 检测证实,oe-circRNA 组 HAS-CIRCpedia-5280 表达水平较 oe-NC 组极显著上调（P<0.01）。对结肠癌裸鼠移植瘤模型开展 hBD-1 腹腔干预试验,相较于 oe-NC+PBS 组,oe-circRNA+PBS 组移植瘤重量极显著增大（P<0.01）、体积显著增大（P<0.05）;oe-NC+hBD-1、oe-circRNA+hBD-1 组移植瘤重量极显著减小（P<0.01）、体积显著减小（P<0.05）。qRT-PCR 检测显示,与oe-NC+PBS 组相比,oe-NC+hBD-1组HAS-CIRCpedia-5280表达水平显著下调（P<0.05）;与oe-circRNA+PBS 组相比,oe-circRNA+hBD-1 组HAS-CIRCpedia-5280 表达显著下调（P<0.05）。[结论] 试验结果表明,HAS-CIRCpedia-5280过表达促进结肠癌移植瘤的生长。进一步研究发现,hBD-1处理可有效下调HAS-CIRCpedia-5280的表达,并抑制移植瘤生长。由此推测hBD-1通过HAS-CIRCpedia-5280抑制结肠癌的进展。

关键词: β-防御素-1, 人β-防御素-1, HAS-CIRCpedia-5280, 结肠癌

Abstract: [Objective] To verify the role of β-defensin-1 in inhibiting the growth of colon cancer via circular RNA HAS-CIRCpedia-5280 by subcutaneous tumorigenesis experiments in nude mice. [Methods] Lentiviral transduction technology was performed to transduce human colon carcinoma HCT-116 cells with the overexpression lentivirus carrying HAS-CIRCpedia-5280 and empty-vector control lentivirus, respectively. After polybrene-assisted infection and puromycin screening for 2 weeks, HAS-CIRCpedia-5280 overexpression stably transduced HCT-116 cell line （HAS-CIRCpedia-5280 cells） and empty vector control stably transduced HCT-116 cell line （NC cells） were constructed. Thirty BALB/c-nu nude mice were divided into 6 groups[oe-NC group, subcutaneously injected with NC cells; oe-circRNA group, subcutaneously injected with HAS-CIRCpedia-5280 cells; oe-NC+PBS group, subcutaneously injected with NC cells and intraperitoneally injected with PBS; oe-circRNA+PBS group, subcutaneously injected with HAS-CIRCpedia-5280 cells and intraperitoneally injected with PBS; oe-NC+hBD-1 （human β-defensin-1） group, subcutaneously injected with NC cells and intraperitoneally injected with hBD-1; oe-circRNA+hBD-1 group, subcutaneously injected with HAS-CIRCpedia-5280 cells and intraperitoneally injected with hBD-1], with 5 mice in each group. The BALB/c-nu nude mice subcutaneous xenograft tumor model was established by subcutaneous injection of the above two kinds of stably transduced cells. The oe-NC+hBD-1 group and the oe-circRNA+hBD-1 group, the oe-NC+PBS group and the oe-circRNA+PBS group were intraperitoneally injected with 0.1 mg/（kg·BW） hBD-1 or an equal volume of PBS, respectively, for intervention, twice a week （Monday and Thursday）, and tumor volume was recorded every 7 days. All injections were completed within 4 weeks from the first administration. The nude mice were euthanized 72 h after the last administration, and the tumor tissues were dissected to measure tumor volume and weight. [Results] HAS-CIRCpedia-5280 overexpression （oe-circRNA） and empty vector control （oe-NC） stably transduced HCT-116 cell lines were constructed by lentivirus transduction. Fluorescence microscopy showed that most cells in the oe-circRNA group and oe-NC group exhibited clear green fluorescence, and the positive rate of cells was higher. The results of qRT-PCR showed that compared with the oe-NC control group, the expression level of HAS-CIRCpedia-5280 in the oe-circRNA group was extremely significantly up-regulated （P<0.01）. A subcutaneous xenograft tumor model of colon cancer in nude mice was constructed, and tumor growth was regularly monitored. The results showed that compared with the oe-NC group, the weight of transplanted tumor in the oe-circRNA group was extremely significantly increased （P<0.01）, and the volume was significantly increased （P<0.05）. qRT-PCR detection confirmed that the expression level of HAS-CIRCpedia-5280 in the oe-circRNA group was extremely significantly up-regulated compared with the oe-NC group （P<0.01）. An hBD-1 intraperitoneal intervention experiment was conducted on the colon cancer xenograft tumor model in nude mice. Compared with the oe-NC+PBS group, the weight of transplanted tumors in the oe-circRNA+PBS group was extremely significantly increased （P<0.01） and the volume was significantly enlarged （P<0.05）; the weight of transplanted tumors in the oe-NC+hBD-1 and oe-circRNA+hBD-1 groups was extremely significantly decreased （P<0.01） and the volume was significantly reduced （P<0.05）. qRT-PCR detection showed that, compared with the oe-NC+PBS group, the expression level of HAS-CIRCpedia-5280 in the oe-NC+hBD-1 group was significantly down-regulated （P<0.05）; compared with the oe-circRNA+PBS group, the expression of HAS-CIRCpedia-5280 in the oe-circRNA+hBD-1 group was significantly down-regulated （P<0.05）. [Conclusion] The experimental results indicate that overexpression of HAS-CIRCpedia-5280 promotes the growth of colon cancer xenograft tumors. Further studies found that hBD-1 treatment can effectively down-regulate the expression of HAS-CIRCpedia-5280 and inhibit the growth of xenograft tumors. It is therefore speculated that hBD-1 inhibits the progression of colon cancer through HAS-CIRCpedia-5280.

Key words: beta-defensin-1, human beta-defensin-1, HAS-CIRCpedia-5280, colon cancer

中图分类号:

S859.79

乔璐, 安世祥, 赵鹏伟, 包丽丽. β-防御素-1通过环状RNA HAS-CIRCpedia-5280抑制小鼠结肠癌的研究[J]. 畜牧与饲料科学, 2026, 47(3): 1-8.

QIAO Lu, AN Shixiang, ZHAO Pengwei, BAO Lili. β-Defensin-1 Inhibits Colon Cancer via Circular RNA HAS-CIRCpedia-5280 in Mice[J]. Animal Husbandry and Feed Science, 2026, 47(3): 1-8.

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