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探索红外光谱监测肿瘤细胞凋亡的可行性及初步分析。采用结肠癌细胞系SW620作为诱导细胞模型,MTT法确定最优5-FU诱导浓度,无血清饥饿培养协同细胞周期阻滞在G1和S期,傅里叶显微红外光谱仪(FTIR)联合流式细胞仪(FCM)对SW 620细胞和凋亡的SW 620细胞12 h、早期凋亡(24 h)和晚期凋亡(48 h)的峰位、相对峰强等指标进行比较分析。光谱分析结果显示SW 620对比凋亡细胞有以下特征:(1)与脂类相关谱带1 740 cm~(-1)相对峰强比I_(1740)/I_(1460)明显降低(p<0.05),表明凋亡细胞中脂类相对含量增多。(2)与氨基酸残基相关谱带1 410 cm~(-1),I_(1460)/I_(1460)在凋亡早期和晚期明显升高(p<0.05),与丝、苏氨基酸相关谱带1 120 cm~(-1)向高波数移动,I_(1120)/I_(1460)明显升高(P<0.05),表明凋亡细胞中DNA双螺旋解链,氨基酸残基增多。(3)DNA的反对称伸缩1 240 cm~(-1)向高波数移动,表明凋亡细胞中核酸分子构象发生改变。(4)与核酸多糖1 040 cm~(-1)相关谱带在凋亡的24和48 h出现,向高波数移动,I_(1040)/I_(1460)在凋亡晚期降低(P<0.05)。初步研究结果表明傅里叶变换红外光谱分析可能成为实时无创监测肿瘤化疗的有效方法。
Feasibility and preliminary analysis of detecting apoptosis of tumor cells by infrared spectroscopy. The colon cancer cell line SW620 was used as an induced cell model. MTT assay was used to determine the optimal concentration of 5-FU. Synergistic cell cycle arrest in serum-free starvation was observed in G1 and S phases. FTIR combined with flow cytometry (FCM), SW 620 cells and apoptosis of SW 620 cells 12 h, early apoptosis (24 h) and late apoptosis (48 h) peak position, relative peak intensity and other indicators were compared. The results of spectral analysis showed that SW 620 had the following characteristics in comparison with apoptotic cells: (1) Relative intensity peak intensity (I 1740) / I 1460 of 1 740 cm -1 was significantly lower than that of lipid related bands (p 0.05 ), Indicating that the relative content of lipids in apoptotic cells increased. (2) The correlation bands (1 410 cm -1, 1460/1460) with amino acid residues were significantly increased in the early and late stages of apoptosis (p <0.05) (1120) / I_ (1460) was significantly increased (P <0.05), indicating that the DNA double helix in the apoptotic cells was melted and the number of amino acid residues increased. (3) The antisymmetric DNA stretching of 1 240 cm ~ (-1) moves toward the higher wave number, indicating that the conformation of nucleic acid molecules in apoptotic cells change. (4) The bands associated with 1 040 cm -1 of nucleic acid polysaccharides appeared at 24 and 48 h after apoptosis, shifted to high wave numbers, and decreased (P <0.05) at I4040 / I1460 ). Preliminary results show that Fourier transform infrared spectroscopy may become a real-time non-invasive monitoring of tumor chemotherapy is an effective method.