Van den Berg E, Van Oven M W, de Jong B, Dam A, Wiersema J, Dijkhuizen T, Hoekstra H J, Molenaar W M
Department of Medical Genetics, University of Groningen, The Netherlands.
Lab Invest. 1994 Mar;70(3):307-13.
Both DNA flow cytometry and cytogenetic analysis have been used to study soft tissue tumors. With flow cytometry, the DNA content of a relatively large number of cells can be examined, but cytogenetic analysis gives more detailed information about genomic changes.
In order to compare the advantages and drawbacks of DNA flow cytometry versus chromosomal analysis, 92 primary or recurrent malignant, 16 borderline malignant, and 13 benign soft tissue tumors were karyotyped after short-term culture. DNA ploidy was determined by flow cytometry of suspensions prepared from frozen or paraffin-embedded samples. From 97 patients, 121 samples were analyzed.
On the basis of the results, four groups were distinguished: DNA-euploid tumors with normal diploid karyotypes (group a) or with abnormal (group b) karyotypes, and DNA-aneuploid tumors with normal (group c) or abnormal (group d) karyotypes. The findings in group b show that structural chromosomal abnormalities or minor numerical aberrations of chromosomes are not detected by DNA flow cytometry. In group c, the finding of tumors with an aneuploid DNA-profile and cells with normal karyotypes is most likely due to overgrowth of fibroblasts during culture and subsequent karyotyping of normal cells.
The findings show that a) DNA flow cytometry has a higher success rate than karyotyping, b) both techniques are complementary, such that DNA flow cytometry gives an "overview", whereas karyotyping gives more detailed information; comparison of both techniques in individual cases leads to a better understanding of the chromosomal events that occurred during oncogenesis, c) histologically low grade tumors are generally DNA-diploid, but may have an abnormal karyotype, and d) histologically high-grade sarcomas tend to have an aneuploid DNA-profile; they are generally more difficult to karyotype.
DNA 流式细胞术和细胞遗传学分析均已用于软组织肿瘤的研究。通过流式细胞术,可以检测相对大量细胞的 DNA 含量,但细胞遗传学分析能提供有关基因组变化的更详细信息。
为比较 DNA 流式细胞术与染色体分析的优缺点,对 92 例原发性或复发性恶性、16 例交界性恶性和 13 例良性软组织肿瘤进行短期培养后进行核型分析。DNA 倍性通过对从冷冻或石蜡包埋样本制备的悬液进行流式细胞术测定。对 97 例患者的 121 个样本进行了分析。
根据结果,区分出四组:具有正常二倍体核型的 DNA 整倍体肿瘤(a 组)或具有异常核型的(b 组),以及具有正常(c 组)或异常(d 组)核型的 DNA 非整倍体肿瘤。b 组的结果表明,DNA 流式细胞术未检测到染色体结构异常或染色体微小数目畸变。在 c 组中,具有非整倍体 DNA 图谱的肿瘤和具有正常核型的细胞的发现很可能是由于培养过程中成纤维细胞过度生长以及随后对正常细胞进行核型分析所致。
结果表明,a)DNA 流式细胞术的成功率高于核型分析,b)两种技术互补,即 DNA 流式细胞术给出“总体情况”,而核型分析提供更详细的信息;在个别病例中比较两种技术有助于更好地理解肿瘤发生过程中发生的染色体事件,c)组织学上低级别肿瘤通常为 DNA 二倍体,但可能具有异常核型,d)组织学上高级别肉瘤往往具有非整倍体 DNA 图谱;它们通常更难进行核型分析。