人卵巢癌动物模型中循环无细胞DNA含量与瘤负荷及细胞凋亡的关系  

Relationship between tumor-related cfDNA and tumor-bearing or cell apoptosis in animal model

作　　者：杨波[1] 吴小华 于利洁[3] 崔洪银[1] 

机构地区：[1]河北医科大学第四医院妇产科,河北石家庄050011 [2]白求恩国际和平医院妇产科,河北石家庄050082 [3]河北医科大学口腔医院,河北石家庄050017

出　　处：《中国现代医学杂志》2010年第16期2459-2463,共5页China Journal of Modern Medicine

基　　金：河北省自然基金资助项目(No:C2005000804)

摘　　要：目的研究循环无细胞DNA(cfDNA)的来源,探讨血清cfDNA水平在卵巢上皮性癌临床诊断及病情监测中的意义。方法利用裸鼠制备人卵巢癌细胞荷瘤模型及腹水模型,并对部分裸鼠实施化疗。应用半定量PCR法检测裸鼠血清及腹水中ALU115、ALU247以及CA125-cfDNA含量,TUNEL法检测肿瘤组织细胞凋亡情况,并且对荷瘤裸鼠化疗前后血清cfDNA水平变化进行检测。结果 cfDNA在各组荷瘤裸鼠的血清及腹水中均有表达,在腹水中的表达值与同一个体血清中含量相比没有显著性差异。血清cfDNA水平与瘤负荷成正相关。cfDNA在化疗后1d达最大值,而后缓慢下降,在化疗10d其值明显低于化疗前水平。化疗前后cfDNA与肿瘤细胞的凋亡率呈正相关。结论血清cfDNA来源于肿瘤细胞,并与细胞凋亡和坏死有关,可能为卵巢上皮性癌的诊断及病情监测提供理论依据。【Objective】To investigate the relationship of the level of cfDNA between tumor-bearing and tumor cells apoptosis,change of cfDNA blood flow kinetics in the serum of female nude mice after human EOC cell line injection.【Methods】The human EOC cell line SKOV3 were injected on the back or intra-abdomen of the 6～8 weeks female nude mice,the xenograft tumor tissues,serum and ascites were collected respectively in different groups.Semi-quantitive RT-PCR was used to analysis CA125-cfDNA,ALU 115 and ALU247 expression level.TUNEL was used to detect cell in-situ apoptosis of the xenograft..The correlation between cfDNA level in the serum or ascites and tumor weight or apoptosis were analyzed.【Results】101 bp fragment of CA125-cfDNA,DNA fragments of ALU 115 and ALU247 were found in the serum or ascites of every groups,especially the 261 bps fragment of CA125-cfDNA level was found in 31 of 78 mice（39.74%）.Serum cfDNA level and tumor-bearing 101 bps fragment of CA125-cfDNA and tumor related cfDNA in serum were positive correlated with tumor weight,the differences were significant in statistics（P 0.05）.101bps fragment of CA125-cfDNA and tumor-related cfDNA in serum reached maximal value on day 1 after chemotherapy,and then slowed down,the values were significant lower on day 10 after chemotherapy than that of pre-chemotherapy.Human specific cfDNA was found in the serum and ascites of tumor-bear nude mice,the cfDNA level increased significantly after chemotherapy and then slowed down,also it correlated with cell apoptosis.101 bps fragment of CA125-cfDNA and tumor related cfDNA are positive correlated with cell apoptosis respectively（P 0.05）.【Conclusion】It was certified that the cfDNA comes from tumor cell and it correlates with apoptosis and necrosis.101 bps fragment of CA125-cfDNA and tumor-related cfDNA in serum positive correlated with tumor-bearing,which shows some useful information for EOC diagnosis and management

关 键 词：卵巢上皮性癌 循环无细胞DNA 半定量PCR 凋亡 

分 类 号：R737.31[医药卫生—肿瘤] R-332[医药卫生—临床医学]