光谱学与光谱分析
光譜學與光譜分析
광보학여광보분석
SPECTROSCOPY AND SPECTRAL ANALYSIS
2013年
12期
3169-3174
,共6页
刘栋%孙学军%张超%贺赛%杜俊凯%霍雄伟%郑见宝%张仕运%张元福%徐怡庄%吴瑾光
劉棟%孫學軍%張超%賀賽%杜俊凱%霍雄偉%鄭見寶%張仕運%張元福%徐怡莊%吳瑾光
류동%손학군%장초%하새%두준개%곽웅위%정견보%장사운%장원복%서이장%오근광
结肠癌%细胞周期%细胞凋亡%红外光谱%化疗
結腸癌%細胞週期%細胞凋亡%紅外光譜%化療
결장암%세포주기%세포조망%홍외광보%화료
Colon cancer%Cell cycle%Apoptosis%FTIR%Chemotherapy
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，M T T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1740 cm-1相对峰强比 I1740/I1460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1410 cm -1， I1460/I1460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1120 cm -1向高波数移动，I1120/I1460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1240 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1040 cm -1相关谱带在凋亡的24和48 h出现，向高波数移动， I1040/I1460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。
探索紅外光譜鑑測腫瘤細胞凋亡的可行性及初步分析。採用結腸癌細胞繫SW620作為誘導細胞模型，M T T法確定最優5-FU誘導濃度，無血清饑餓培養協同細胞週期阻滯在G1和S期，傅裏葉顯微紅外光譜儀（FTIR）聯閤流式細胞儀（FCM ）對SW 620細胞和凋亡的SW 620細胞12 h、早期凋亡（24 h）和晚期凋亡（48 h）的峰位、相對峰彊等指標進行比較分析。光譜分析結果顯示SW 620對比凋亡細胞有以下特徵：（1）與脂類相關譜帶1740 cm-1相對峰彊比 I1740/I1460明顯降低（ p＜0.05），錶明凋亡細胞中脂類相對含量增多。（2）與氨基痠殘基相關譜帶1410 cm -1， I1460/I1460在凋亡早期和晚期明顯升高（ p＜0.05），與絲、囌氨基痠相關譜帶1120 cm -1嚮高波數移動，I1120/I1460明顯升高（p＜0.05），錶明凋亡細胞中DNA雙螺鏇解鏈，氨基痠殘基增多。（3）DNA的反對稱伸縮1240 cm-1嚮高波數移動，錶明凋亡細胞中覈痠分子構象髮生改變。（4）與覈痠多糖1040 cm -1相關譜帶在凋亡的24和48 h齣現，嚮高波數移動， I1040/I1460在凋亡晚期降低（ p＜0.05）。初步研究結果錶明傅裏葉變換紅外光譜分析可能成為實時無創鑑測腫瘤化療的有效方法。
탐색홍외광보감측종류세포조망적가행성급초보분석。채용결장암세포계SW620작위유도세포모형，M T T법학정최우5-FU유도농도，무혈청기아배양협동세포주기조체재G1화S기，부리협현미홍외광보의（FTIR）연합류식세포의（FCM ）대SW 620세포화조망적SW 620세포12 h、조기조망（24 h）화만기조망（48 h）적봉위、상대봉강등지표진행비교분석。광보분석결과현시SW 620대비조망세포유이하특정：（1）여지류상관보대1740 cm-1상대봉강비 I1740/I1460명현강저（ p＜0.05），표명조망세포중지류상대함량증다。（2）여안기산잔기상관보대1410 cm -1， I1460/I1460재조망조기화만기명현승고（ p＜0.05），여사、소안기산상관보대1120 cm -1향고파수이동，I1120/I1460명현승고（p＜0.05），표명조망세포중DNA쌍라선해련，안기산잔기증다。（3）DNA적반대칭신축1240 cm-1향고파수이동，표명조망세포중핵산분자구상발생개변。（4）여핵산다당1040 cm -1상관보대재조망적24화48 h출현，향고파수이동， I1040/I1460재조망만기강저（ p＜0.05）。초보연구결과표명부리협변환홍외광보분석가능성위실시무창감측종류화료적유효방법。
The aim of the present study was to evaluate Fourier transform infrared spectroscopy (FTIR) monitoring of biochemi-cal changes in apoptosis cells .Different concentrations of 5-fluorouracil (5-FU) treated colon cancer cell lines SW620 were used to determine the optimum concentration of 5-FU IC50 by means of MTT assay .Cell starvation and 5-Fu synergistic cell cycle ar-rest was in G1 and S phase .FTIR combined with flow cytometry was applied to analysis of SW 620 cells and SW620 cells treated with 5-FU for 12h ,24h (early apoptosis) and 48 h (late apoptosis) respectively .The peak position and the intensity of all bands were measured and comparison was made between the SW 620 and apoptotic SW620 cells .Apoptosis cells have following charac-teristics compared with SW620 cells (1) The band at 1 740 cm-1 is an CO stretching vibration .Changes in these bands can reflect lipid changes ,and relative peak intensity ratio I1 740/I1 460 significantly increased (p<0.05) ,indicating that the relative contents of lipid in apoptosis cells increased .(2) The band at the 1 410 cm -1 peak represents that C-H stretching related was increased to amino acid residues and shifted to higher wave numbers compared to other groups .I1 410/I1 460 at early and late death phase was significantly increased ,which suggests that the relative contents of amino acid residues in apoptosis cells increased (p<0.05) .New vibrational bands at 1 120 cm-1 appeared at 24h and increased at 48 h compared with other groups .The 1 120 cm -1 absorption band is mainly due to ser ,serine and threonine C-O(H) stretching vibration ,and I1 120/I1 460 significantly in-creased (p<0.05) ,indicating that the relative quantity of amino acid residues in apoptosis cells increased due to that DNA un-winds the double helix .(3) 1 240 cm -1 is mainly due to the asymmetric stretching modes of phosphodiester groups shifting to higher wave number ,illustrating that nucleic acid conformation was changed in apoptosis cells .(4) The band 1 040 cm -1 associ-ated with polysaccharide appeared at 24 and 48 h ,meanwhile shifted to higher wave number ,suggesting that polysaccharide de-creased in late apoptotic cells ,and I1 040/I1 460 increased at late stage apoptosis ,indicating that the relative content of polysaccha-ride in apoptosis cells increased .The authors’ results suggest that FTIR applied to monitoring SW 620 cells apoptosis may be as a potential diagnostic tool for cancer chemotherapy monitoring .
