结直肠癌中葡萄糖转运蛋白-12作为化疗联合用药靶点的研究

GLUT 12 in colorectal cancer as a target for combination therapy

目的研究葡萄糖转运蛋白-12(GLUT12)在结直肠癌(CRC)细胞和组织中的表达与功能,探讨其作为联合用药治疗靶点的可能性。方法分别采用RT-PCR和蛋白免疫印迹法检测CRC及正常结肠中GLUT12基因和蛋白水平。通过荧光免疫及蛋白印迹法观察槲皮素(QC)对肿瘤细胞中GLUT12定位的调节作用。四氮唑(MTT)比色法和克隆形成实验(CFA)观察QC和奥沙利铂(OXA)联合使用对CRC细胞增殖的影响。结果 GLUT12基因在肿瘤组织中的相对表达水平高于正常组织[(3. 2±0. 3) vs(0. 5±0. 6),P <0. 05]。高糖培养液能改变GLUT12在细胞中的定位及表达水平,QC能抑制高糖对GLUT12蛋白的调节作用。MTT和CFA试验显示,OXA和QC联用能使CRC细胞的活力降低[OXA+OC:(63. 3±7. 1)%vs OC:(88. 7±4. 2)%,P <0. 05]。QC和OXA联用能抑制结肠癌细胞群落的增殖能力。结论 QC能有效抑制CRC细胞GLUT12蛋白的功能,能增加OXA杀伤CRC的作用。

Objective To study the expression and function of GLUT12 in CRC cells and tissues, andto assess the possibility of se-lecting it as a therapeutic target for combination therapy. Methods The gene and protein status of GLUT12 in CRC and the normal colon were examined using RT - PCR and Western blot methods. The regulatory effect of OC on the cellular localization and protein expression of GLUT12 were evaluated using immunocytochemistry and Western blotting. We then studied the synergistic effect of QC in combination with OXA against CRC cells using MTT and CFA assays. Results The protein level of GLUT12 in CRC tissues was higher in comparison with the normal value （3.2±0.3 vs 0.5 ±0.6, 05）. High glucose - containing media could alter the localization and expression levels ofGLUT12 in CRC cells, which could be blocked by QC. MTT tests showed that OXA in combination with QC could significantly reduce the vi- ability of CRC cells（OXA ＋ QC 63.3 ±7.1% QC： 88.7 ±4.2% , P 〈 0 . 0 5 ） . The addition of QC into OXA could significantly suppress the viability of CRC cells. Conclusion QC can effectively inhibit the function of significantly higher level of GLUT12 in CRC cells, and QC could enhance the ability of OXA to kill CRC cells.