目的 探讨缺氧诱导因子抑制剂YC-1对口腔鳞癌(OSCC)中葡萄糖转运蛋白-1(Glut1)、血管内皮生长因子(VEGF)基因的调控作用。方法 (1)应用免疫组织化学法检测60例OSCC患者和12例癌旁正常组织中的缺氧诱导因子-1α(HIF-1α)、Glut1、VEGF的表达。(2)人舌癌Tca8113细胞在体外进行培养,将其分成缺氧组、常氧组。将浓度为0、10、100μmmol/L的HIF-1α抑制剂YC-1分别加入二组后,通过RT-PCR技术检测OSCC细胞中HIF-1α、Glut1、VEGF mRNA的转录水平。结果 免疫组化发现:OSCC癌中HIF-1α、Glut1、VEGF的阳性表达明显多于正常组织,其中HIF-1α的阳性表达与Glut1的阳性表达、VEGF的阳性表达均呈正相关。RT-PCR结果表明,加入YC-1后,HIF-1α的表达在缺氧组和常氧组中均降低,差异有统计学意义(P <0. 05)。HIF-1α的表达在缺氧组随着YC-1浓度不同无剂量依从性,差异无统计学意义(P> 0. 05)。在YC-1作用下,缺氧组和对照组Glut1、VEGF在人舌癌Tca8113细胞中的表达均显著(P <0. 05)降低,缺氧组内Glut1、VEGF的表达随着YC-1浓度增加有剂量依从性(P <0. 05)。结论 HIF-1α、Glut1、VEGF高表达于OSCC组织中。YC-1显著抑制人舌癌Tca8113细胞HIF-1α、Glut1、VEGF分子的表达;YC-1可能通过下调HIF-1α、Glut1、VEGF的表达来抑制OSCC的生长。
Objective To study the function and mechanism of YC-1 on activity of Glut1 and VEGF genes in Oral squamous cell carcinoma. Methods ( 1) HIF-1α,Glut1 and VEGF proteins were detected by immunohistochemical methods in 60 cases of oral squamous cell carcinoma and 12 cases of normal tissue adjacent to tumor. ( 2) Human tongue squamous cancer cell line Tca8113 were incubated under hypoxic or normoxia conditions and were divided into the hypoxia group and the normoxic group. Different concentrations of YC-1 ( 0,10,and 100 μmol /L) were added to both groups.RT-PCR was used to detect the HIF-1α,Glut1 and VEGF mRNA. Results The positive expression rate of HIF-1α,Glut1 and VEGF in OSCC was higher than that in normal tissue.Positive correlation was found between HIF-1α and Glut1,VEGF expression ( P < 0. 05) .RT-PCR results showed that after the adding of YC-1,the expression of HIF-1α in both groups decreased,with statistical significant difference ( P < 0. 05) . With different YC-1 concentration,the expression of HIF-1α in the hypoxia group had no dose dependence,with no statistical significant difference ( P > 0. 05) . Glut1 and VEGF molecular expressions in human tongue squamous cancer cell line Tca8113 decreased significantly in different YC-1 concentrations ( P < 0. 05) . The expressions of Glut1 and VEGF had dose dependence in the hypoxia group with the increase of YC-1 concentrations ( P < 0. 05) . Conclusion The expression levels of HIF-1α,Glut1 and VEGF in oral squamous cell carcinoma are higher than those in the normal tissues. YC-1 inhibits HIF-1α,Glut1 and VEGF expressions in Tongue squamous cancer cell line Tca8113. YC-1 inhibits the growth of OSCC possibly through down-regulating the expressions of HIF-1α,Glut1 and VEGF.
Journal of Binzhou Medical College