沉默AQP3基因表达对结直肠癌细胞增殖、凋亡和化疗敏感性的影响

郑斯鑫 周春华 孔文成 尹光

[摘要] 目的 探討沉默水通道蛋白3（AQP3）基因表达对结直肠癌细胞增殖、凋亡和化疗敏感性的影响。 方法 体外培养结直肠癌细胞LoVo，采用梯度浓度诱导法建立紫杉醇耐药细胞株LoVo/Taxol。将LoVo细胞和LoVo/Taxol细胞分别分为空白对照组（细胞不做任何特殊处理）、阴性对照组（转染阴性对照质粒）、shAQP3组（沉默AQP3表达），逆转录-聚合酶链反应（qRT-PCR）和Western blot法检测AQP3 mRNA和蛋白表达。CCK-8实验检测细胞的增殖能力，流式细胞术检测细胞的凋亡情况。四唑化合物（MTS）检测紫杉醇的细胞毒性。结果 人结肠癌细胞LoVo中AQP3 mRNA相对表达量和蛋白表达量均高于人正常结肠上皮细胞NCM460和HCoEpiC，差异有统计学意义（P<0.05）。敲低AQP3表达后，shAQP3组LoVo细胞增殖活性（OD值）较空白对照组和阴性对照组降低，同时凋亡率升高，差异有统计学意义（P<0.05）。成功构建LoVo/Taxol耐药细胞，对紫杉醇、5-Fu、阿霉素的耐药指数分别为（31.97±1.78）、（17.97±2.14）、（104.80±5.53）。而shAQP3组LoVo/Taxol细胞对紫杉醇、5-Fu、阿霉素的耐药指数分别为（6.06±0.25）、（5.57±0.20）、（21.23±0.27），较空白对照组和阴性对照组明显降低，差异有统计学意义（P<0.05）。 结论 沉默AQP3基因表达能够抑制结直肠癌细胞增殖，促进结直肠癌细胞凋亡并增强LoVo/Taxol耐药细胞的化疗敏感性。

[关键词] AQP3;结直肠癌;增殖;凋亡;化疗敏感性

[中图分类号] R735.3          [文献标识码] A          [文章编号] 1673-9701（2021）31-0006-06

[Abstract] Objective To explore the effect of silencing aquaporin 3 （AQP3） gene expression on the proliferation， apoptosis， and chemosensitivity of colorectal cancer cells. Methods LoVo colorectal cancer cells were cultured in vitro， and the LoVo/Taxol paclitaxel-resistant cell lines were established by gradient concentration induction method. The LoVo cells and LoVo/Taxol cells were divided into the blank control group （no special treatment）， the negative control group （negative control plasmid transfection）， and the shAQP3 group （silencing AQP3 expression）. Reverse transcription-polymerase chain reaction （qRT-PCR） and Western blot method were used to detect AQP3 mRNA and protein expression. CCK-8 experiment was used to detect cell proliferation ability， and flow cytometry was used to detect cell apoptosis. Tetrazolium compound （MTS） was used to detect the cytotoxicity of paclitaxel. Results The AQP3 mRNA relative expression and protein expression in human colon cancer cells LoVo were higher than those in human normal colonic epithelial cells NCM460 and HCoEpiC，the difference was statistically significant （P<0.05）. After knocking down the AQP3 expression， the proliferation activity （OD value） of LoVo cells in the shAQP3 group was lower than those in the blank control group and the negative control group， and the apoptosis rate was higher than those in the blank control group and the negative control group，the difference was statistically significant（P<0.05）. The LoVo/Taxol resistant cells were successfully constructed， and the resistance indexes to paclitaxel， 5-Fu and adriamycin were （31.97±1.78）， （17.97±2.14）， （104.80±5.53）， respectively. The resistance indexes of LoVo/Taxol cells to paclitaxel， 5-Fu and adriamycin in the shAQP3 group were （6.06±0.25）， （5.57±0.20）， （21.23±0.27）， respectively， which were significantly lower than those in the blank control group and the negative control group，the difference was statistically significant（P<0.05）. Conclusion Silencing AQP3 gene expression can inhibit the proliferation of colorectal cancer cells， promote the apoptosis of colorectal cancer cells， and enhance the chemosensitivity of LoVo/Taxol resistant cells.

[Key words] AQP3; Colorectal cancer; Proliferation; Apoptosis; Chemosensitivity

2018年，中国结直肠癌发病率占世界的24.3%，病死率占世界的22.9%，且逐渐呈现年轻化趋势[1]。为了提高临床治疗效果，有必要更好地了解结直肠癌的发生机制，从而推动精准医学的发展。水通道蛋白（Aquaporins，AQPs）是一种糖基化的膜整合蛋白，能够通过跨质膜的渗透梯度促进跨上皮水分子转运[2]。……