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结直肠癌中5′tiRNA-Val-CAC对增殖的影响及病理意义

Effect of 5′tiRNA-Val-CAC on proliferation and its pathological significance in colorectal cancer

    摘要
  • 摘要:
    目的 探讨5′tiRNA-Val-CAC在结直肠癌(CRC)中的表达、临床病理意义及其对CRC细胞生长与增殖的影响。
    方法 分析转运RNA衍生的小RNA(TDSR)表达谱，并通过实时荧光定量聚合酶链反应(qRT-PCR)检测5′tiRNA-Val-CAC表达水平。采用RNA原位杂交技术(RISH)检测82例CRC患者CRC组织及癌旁正常组织中5′tiRNA-Val-CAC阳性表达情况，并分析5′tiRNA-Val-CAC阳性表达与患者临床病理特征的相关性。通过CCK-8实验和细胞克隆平板实验检测5′tiRNA-Val-CAC对CRC细胞生长和增殖的影响。
    结果 qRT-PCR结果显示, CRC组织中5′tiRNA-Val-CAC表达水平高于癌旁正常组织，差异有统计学意义(t=9.111, P=0.012); CRC细胞系SW620、HCT116中5′tiRNA-Val-CAC表达水平高于正常人肠上皮细胞(HIEC), 差异有统计学意义(t=11.475、7.158, P=0.008、0.019)。RISH实验结果显示, CRC组织中5′tiRNA-Val-CAC阳性表达率为82.93%(68/82), 而癌旁正常组织中阳性表达率为7.32%(6/82), 差异有统计学意义(χ2=31.444, P < 0.001)。5′tiRNA-Val-CAC高阳性表达与CRC患者肿瘤直径(χ2=6.924, P < 0.05)和分化程度(χ2=25.030, P < 0.05)具有相关性，但与性别、年龄、发病部位、浸润深度、淋巴结转移和TNM分期无相关性(P>0.05)。CCK-8实验和细胞克隆平板实验结果显示，与对照细胞比较，过表达5′tiRNA-Val-CAC的CRC细胞增殖活性升高、细胞克隆数目增多，而敲低5′tiRNA-Val-CAC表达的CRC细胞增殖活性下降、细胞克隆数目减少，差异均有统计学意义(P < 0.05)。
    结论 5′tiRNA-Val-CAC在CRC组织中高表达，能促进CRC细胞的增殖与生长，影响CRC的进展和预后。5′tiRNA-Val-CAC或可成为CRC增殖的分子标志物，用于CRC的诊断和预后判断。

     

    Abstract:
    Objective To investigate the expression of 5′tiRNA-Val-CAC in colorectal cancer (CRC), its clinicopathological significance and its effect on tumor growth and proliferation.
    Methods The expression profile of tRNA derived small RNA (TDSR) was analyzed and the expression level of 5′tiRNA-Val-CAC was detected by quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR). RNA in situ hybridization (RISH) was used to detect the positive expression of 5′tiRNA-Val-CAC in CRC tissues and normal adjacent tissues of 82 CRC patients. The correlation between the positive expression and clinicopathological features of patients was statistically analyzed. CCK-8 experiment and cell cloning plate experiment were used to observe the effect of 5′tiRNA-Val-CAC on the growth and proliferation of colorectal cancer cells.
    Results qRT-PCR results showed that the expression level of 5′tiRNA-Val-CAC in CRC tissues was significantly higher than that in adjacent normal colorectal tissues(t=9.111, P=0.012); the expression levels of 5′tiRNA-Val-CAC in CRC cell lines SW620 and HCT116 were higher than those in normal intestinal epithelial cells (HIEC), the difference was statistically significant (t=11.475, 7.158, P=0.008, 0.019). RISH experimental results showed that positive rate of 5′tiRNA-Val-CAC expression in CRC tissues was 82.93%(68/82), and was 7.32%(6/82) in adjacent normal tissues, which showed a significant differences(χ2=31.444, P < 0.001). Highly positive expression of 5′tiRNA-Val-CAC was correlated with tumor size(χ2=6.924, P < 0.05) and tumor differentiation(χ2=25.030, P < 0.05), but had no correlations with gender, age, lesion location, depth of invasion, lymph node metastasis and TNM stage (P>0.05). CCK-8 experiment and cell cloning plate experiment showed that the proliferation activity of overexpressed 5′tiRNA-Val-CAC colorectal cancer cells and the number of cell clones increased, while proliferation activity of CRC cells knockdown of 5′tiRNA-Val-CAC expression and the number of cell clones decreased compared with the control group(P < 0.05).
    Conclusion 5′tiRNA-Val-CAC that is highly expressed in CRC tissues can promote the proliferation and growth of CRC cells and affect the progression and prognosis of CRC. 5′tiRNA-Val-CAC may be a molecular marker of CRC proliferation and can be used as an index for diagnosis and prognosis of CRC.

     

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