Pathak R, Sarma A, Sengupta B, Dey S K, Khuda-Bukhsh A R
Department of Biotechnology, West Bengal University of Technology, Salt Lake, Kolkata.
Int J Radiat Biol. 2007 Jan;83(1):53-63. doi: 10.1080/09553000601085964.
To study the effects of 12C-beam of 295 keV/microm (57.24 MeV) on M5 and Chinese hamster V79 cells by using cytogenetic assays like micronuclei (MN) induction, chromosomal aberrations (CA) and apoptosis. Additionally, the relative survival of these two cell lines was tested by the colony forming ability of the cells, with a view to understanding the mechanism of cellular damages that lead to difference in cell survival.
Confluent cells were irradiated with 12C-beam at various doses using 15UD Pelletron accelerator. Cell survival was studied by the colony forming ability of cells. MN assay was done by fluorescent staining. Different types of chromosomal aberrations in metaphase cells were scored at 12 h after irradiation. Apoptosis was measured at different post irradiation times as detected by nuclear fragmentation and DNA ladder was prepared after 48 h of incubation.
Dose-dependent decrease in surviving fractions was found in both the cell lines. However, the surviving fractions were higher in M5 cells in comparison to V79 cells when exposed to the same radiation doses. On the other hand, induced MN frequencies, CA frequencies and apoptosis percentages were less in M5 cells than V79 cells. Very good correlations between surviving fractions and induced MN frequencies or induced total CA or induced apoptosis percentages were obtained in this study.
The cell strain M5 showed relatively more radio-resistance to 12C-beam compared to Chinese hamster V79 cells in this study. As the MN formation, CA and apoptosis induction were less in M5 cells as compared to parental V79 cells, the higher cell survival in the former could possibly be attributed to their better repairing ability leading to higher cell survival.
通过微核(MN)诱导、染色体畸变(CA)和凋亡等细胞遗传学检测方法,研究295 keV/μm(57.24 MeV)的12C束对M5细胞和中国仓鼠V79细胞的影响。此外,通过细胞集落形成能力检测这两种细胞系的相对存活率,以了解导致细胞存活差异的细胞损伤机制。
使用15UD Pelletron加速器,以不同剂量的12C束照射汇合的细胞。通过细胞集落形成能力研究细胞存活情况。通过荧光染色进行微核检测。在照射后12小时对中期细胞中不同类型的染色体畸变进行评分。在照射后的不同时间测量凋亡情况,通过核碎片化检测,孵育48小时后制备DNA梯带。
在两种细胞系中均发现存活分数呈剂量依赖性下降。然而,当暴露于相同辐射剂量时,M5细胞的存活分数高于V79细胞。另一方面,M5细胞中诱导的微核频率、染色体畸变频率和凋亡百分比均低于V79细胞。本研究中获得了存活分数与诱导的微核频率、诱导的总染色体畸变或诱导的凋亡百分比之间非常好的相关性。
在本研究中,与中国仓鼠V79细胞相比,细胞系M5对12C束显示出相对更高的放射抗性。由于与亲本V79细胞相比,M5细胞中的微核形成、染色体畸变和凋亡诱导较少,前者较高的细胞存活率可能归因于其更好的修复能力,从而导致更高的细胞存活率。