模拟微重力条件HN13细胞的生长特点

Cultivation of oral squamous cell carcinoma cells in a simulated microgravity environment

目的:观察口腔鳞状细胞癌细胞HN13在模拟微重力条件下的形态学特点和生长情况,检测肿瘤细胞分化和侵袭相关基因的表达,并分析其生物学意义。方法:将HN13细胞接种于聚乳酸-羟基乙酸(poly lactic-co-glycolicacid,PLGA),应用旋转细胞培养系统(rotary cell culture system,RCCS)进行模拟微重力培养14d,光学倒置显微镜下观察细胞的形态,以实时定量PCR检测分析立体培养和平面培养细胞中,HIF1α、TGFβ1、VEGF-C、NF-κB和CCND1等基因的表达。采用SPSS17.0软件包对数据进行统计学分析。结果:细胞贴附于PLGA材料表面呈立体生长,表现为典型角化细胞形态,并形成类角化珠样结构,与口腔鳞状细胞癌细胞形态相似。HIF1α,TGFβ1和NF-κB表达降低(P<0.05),VEGF-C表达升高(P<0.05),CCND1表达无显著改变(P>0.05)。结论:模拟微重力条件下三维培养的细胞更接近体内细胞的真实形态,模拟微重力可作为构建体外细胞三维培养模型的方法。

PURPOSE：To study the morphological changes and the different expression of 6 related genes in oral squamous cell carcinoma（OSCC） cells cultured in simulated microgravity（SM） and static condition.METHODS：About 1×108 HN13 cells were seeded into the rotary cell culture system（RCCS） with poly lactic-co-glycolic acid（PLGA）.After cultured for 14 days,the morphological features were observed under phase-contrast microscope.mRNA levels of HIF1α,TGFβ1,VEGF-C,NF-кB and CCND1 between cells cultured in the two conditions were also examined by quantitative RT-PCR assay.All statistical analyses were performed using SPSS 17.0 software package.RESULTS： HN13 cells cultured in RCCS for 14 days were adhered to the PLGA scaffolds and grew in a three-dimensional pattern.The multicellular spheroids in the three-dimensional culture showed a histological pattern similar to the histology observed in human primary OSCC.The expression levels of OSCC-related genes,including HIF1α,TGFβ1 and NF-κB,were significantly lower in cells from the static condition（P0.05）whereas VEGF-C expression level was significantly higher（P0.05）.No difference was observed for CCND1 mRNA level between the cells cultured in the two different conditions.CONCLUSIONS： Establishing SM model of OSCC cell lines may be a prospective method for studying the molecular and biological mechanisms of OSCC and for testing novel target genes for cancer therapy.Supported by Shanghai Leading Academic Discipline Project（S30206）.