| 姬宏利,姬宏娟,王保健.肺癌线粒体 DNA诱导小鼠胚胎成纤维细胞 NIH3T3恶性转化的研究[J].安徽医药,2021,25(3):533-536. |
| 肺癌线粒体 DNA诱导小鼠胚胎成纤维细胞 NIH3T3恶性转化的研究 |
| TheresearchofmitochondrialDNA in lung cancercellon inducingthemutationin NIH3T3 cells |
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| DOI：10.3969/j.issn.1009-6469.2021.03.025 |
| 中文关键词: 肺肿瘤 DNA，线粒体 成纤维细胞 突变 整合 |
| 英文关键词: Lung neoplasms DNA, mitochondrial Fibroblasts Mutation Integration |
| 基金项目:解放军联勤保障部队第九八八医院(原一五三医院)科研基金( 2012013) |
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| 中文摘要: |
| 目的观察转染了 A549及 H226肺癌细胞线粒体 DNA(mtDNA)的小鼠胚胎成纤维细胞 NIH3T3的生物学行为的变化。方法将重组载体 pcDNA3.1(+)-A549 mtDNA、pcDNA3.1(+)-H226 mtDNA通过 Lipofection2000TM转染 NIH3T3细胞；荧光原位杂交( FISH)法观察 mtDNA在核内的整合情况；分析转染前后细胞染色体核型；流式细胞仪( FCM)检测转染细胞的凋亡；四甲基偶氮唑盐微量酶反应比色法( MTT法)测定不同组别细胞的增殖率。结果转染 A549-mtDNA、H226-mtDNA的 NIH3T3细胞染色体出现了易位、断裂的结构畸形，其畸形的多倍体数目百分比( 11.78±0.60)%、(11.33±1.05)%和染色体数目( 3.55±0.57)%、(5.02±0.72)%高于未转染的 NIH3T3细胞( 1.38±0.17)%、(0.54±0.10)%。荧光显微镜观察到 mtDNA能够整合在转染 A549-mtDNA、H226-mtDNA的 NIH3T3细胞染色体间期核中。同时，转染了 A549-mtDNA、H226-mtDNA重组表达载体， A549mtDNA组( 5.20±0.20)%、H226-mtDNA组( 7.75±0.11)%较 NIH3T3组( 19.04±0.08)%细胞的凋亡率明显下降( F=8 381.21，P<0.001)。分别培养 24 h、48 h、72 h，对 MTT的吸光度， NIH3T3/A549-mtDNA组( 0.58±0.06)、(0.96±0.08)、(1.45±0.06)，NIH3T3/ H226-mtDNA组( 0.64±0.04)、(1.02±0.06)、(1.52±0.03)高于空 NIH3T3组( 0.26±0.04)、(0.42±0.03)、(0.81±0.04)、(1.18±0.09)。结论突变的肺癌 mtDNA通过在核内的整合从而影响 NIH3T3细胞的染色体核型及细胞增殖、凋亡等生物学行为。 |
| 英文摘要: |
| Objective To observe the changes of biological behavior of NIH3T3 transfected with mitochondrial DNA (mtDNA) ofA549 and H226 lung cancer cells. Methods The mtDNA eukaryotic expression vector of lung cancer cell was transfected intoNIH3T3 cells by liposome method (Lipfection2000TM). The integration of mtDNA in nucleus was observed by fluorescence in situ hy?bridization (FISH). The pro-and post-transfection chromosomal nucleus cells were analyzed. The apoptosis of NIH3T3 in differentgroup were detected by flow cytometry (FCM). The proliferation rate of different groups NIH3T3 are also determined by MTT method.Results After NIH3T3 cells transfected with A549 mtDNA and h226 mtDNA ,the percentage of abnormal polyploid number (11.78 ±0.60), (11.33 ± 1.05) and chromosome number (3.55 ± 0.57), (5.02 ± 0.72) were higher than those of untransfected NIH3T3 cells (1.38± 0.17), (0.54 ± 0.10).We found that mtDNA could be integrated into interphase nuclei of NIH3T3 cells transfected with A549 mtDNAand h226 mtDNA by Fluorescence microscopy. Moreover, The apoptotic rate in A549-mtDNA group (5.20 ± 0.20)% and H226-mtDNA group (7.75 ± 0.11)% was significantly lower than NIH3T3 group (19.04 ± 0.08)% (F = 8 381.21, P < 0.001). After the different group NIH3T3 were culture for 24 h, 48 h, 72 h, We found that the NIH3T3/A549-mtDNA group absorbance was (0.58±0.06),(0.96±0.08), (1.45±0.06), the NIH3T3/H-226 mtDNA group absorbance was (0.64 ± 0.04), (1.02 ± 0.06), (1.52 ± 0.03), They were higher than inNIH3T3 group (0.42 ± 0.03), (0.81 ± 0.04), (1.18 ± 0.09). Conclusion The mutant-lung cancer mtDNA can affect the karyotype, cellproliferation, apoptosis and other biological behaviors of NIH3T3 cells through nuclear integration. |
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