| 孙奋琪,曹小梅,邹佳杰.肌醇多聚磷酸-4-磷酸酶Ⅱ型表达对结肠癌预后及5-FU化疗敏感性的作用研究[J].中国肿瘤,2022,31(4):311-320. |
| 肌醇多聚磷酸-4-磷酸酶Ⅱ型表达对结肠癌预后及5-FU化疗敏感性的作用研究 |
| Relationship of INPP4B Expression with Prognosis of Colon Cancer and Its Effect on Chemosensitivity of Colon Cancer Cells |
| 投稿时间：2022-01-09 |
| DOI：10.11735/j.issn.1004-0242.2022.04.A009 |
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| 中文关键词: 肌醇多聚磷酸-4-磷酸酶Ⅱ型 结肠癌 氟尿嘧啶 细胞周期 细胞凋亡 |
| 英文关键词:INPP4B colon cancer 5-FU cell cycle apoptosis |
| 基金项目:国家自然科学基金（81972840） |
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| 中文摘要: |
| 摘 要：［目的］ 研究肌醇多聚磷酸-4-磷酸酶Ⅱ型(inositol polyphosphate 4-phosphatase type Ⅱ，INPP4B)的表达对结肠癌预后及氟尿嘧啶(5- fluorouracil，5-FU) 药物敏感性的影响及其机制。［方法］ 通过免疫组织化学方法和R2在线工具分析INPP4B在结肠癌组织的表达及其与预后的关系。构建敲减或过表达INPP4B质粒，分别转染结肠癌细胞株WiDr细胞和SW480细胞，通过实时荧光定量PCR技术检测INPP4B mRNA的表达情况。通过Western blot技术检测细胞中INPP4B、磷酸化的Akt（pAkt）和p27的表达情况。使用CCK-8检测细胞的增殖能力。采取流式细胞术检测细胞的凋亡率和细胞周期的分布。 ［结果］ INPP4B高表达与结肠癌患者预后不良相关（P<0.05）。 敲减INPP4B后，5-FU处理细胞24 h，发现与WiDr shControl细胞相比，WiDr shINPP4B细胞存活率显著降低（P=0.011），凋亡率显著增高（P<0.001），G0/G1期细胞比例显著升高（P<0.001）。过表达INPP4B后，与SW480空载细胞相比，SW480 INPP4B细胞存活率显著升高（P=0.002），凋亡率显著降低（P=0.002），G0/G1期的细胞比例显著降低（P=0.001）。5-FU处理结肠癌细胞后，与WiDr shControl细胞相比，WiDr shINPP4B细胞中pAkt (Ser473)的水平显著降低（P=0.003），pAkt (Thr308)的水平显著降低（P<0.001），p27蛋白的表达显著升高（P<0.001）；与SW480空载细胞相比，SW480 INPP4B细胞中pAkt (Ser473)的水平显著升高（P<0.001），pAkt (Thr308)的水平显著升高（P<0.001)，p27蛋白的表达显著降低（P<0.001）。［结论］ INPP4B高表达与结肠癌患者不良预后相关，并通过激活Akt/p27信号轴降低结肠癌细胞对5-FU的敏感性。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the relationship of inositol polyphosphate 4-phosphatase type Ⅱ(INPP4B) with prognosis of colon cancer and its effect on chemosensitivity of colon cancer cells. ［Methods］ The correlation between expression of INPP4B and prognosis of colon cancer patients was analyzed with bioinformatics methods. Lentivirus-mediated knock-down or overexpressing INPP4B cell lines were constructed with human colon cancer WiDr or SW480 cells. The expression of NPP4B mRNA and protein was detected by real-time quantitative PCR(qPCR) and Western blotting， respectively； the phosphorylated Akt(pAkt) and p27 were detected by Western blotting； cell proliferation ability was measured with cytotoxicity kit-8(CCK-8)； the apoptotic rate and cell cycle distribution were determined with flow cytometry. ［Results］ Kaplan-Meier survival analysis showed that the high expression of INPP4B was correlated with poor prognosis of colon cancer patients(P<0.05). Compared to control cells， INPP4B knockdown increased the sensitivity of colon cancer cells to 5-FU significantly(P=0.011)， enhancing 5-FU-induced cell apoptosis(P<0.001) and increasing 5-FU-induced G1 arrest(P<0.001) of colon cancer cells. Compared to vector control， over-expression of INPP4B decreased the sensitivity of colon cancer cells to 5-FU significantly(P=0.002)， attenuated 5-FU-induced cell apoptosis(P=0.002) and decreasing 5-FU-induced G1 arrest(P=0.001) of colon cancer cells. Knockdown of INPP4B in colon cancer cells significantly decreased p-AKT protein expression(P=0.003) and enhanced p27 protein expression(P<0.001) compared to control cells. Overexpression of INPP4B in colon cancer cells increased p-AKT protein expression(P<0.001) and reduced p27 protein expression(P<0.001). ［Conclusion］ INPP4B is correlated with poor prognosis of colon cancer patients and it can decrease the sensitivity of colon cancer cells to 5-FU via AKT/ p27 signal pathway. |
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