Vihko K K, Seppänen M, Henttinen T, Punnonen J, Grénman S, Punnonen R
Medical School, Department of Obstetrics and Gynecology, University of Tampere, Finland.
Cancer Immunol Immunother. 1997 Jan;43(6):368-74. doi: 10.1007/s002620050346.
The biology and pathogenesis of vulvar carcinoma are poorly understood at present. In order to understand this disease better, we have used recently developed squamous cell carcinoma lines of the vulva as models. Two cell lines originating from two individuals (UM-SCV-1A and UM-SCV-6) were cultured in vitro in 10% fetal calf serum. The effects of interleukins 10 and 13, interferons alpha and gamma, granulocyte/macrophage-growth-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF alpha), and transforming growth factor beta (TGFbeta) on the proliferation of the cells was investigated by using radioactively labelled uridine as tracer. In addition, an investigation on the molecular structure of extracted cellular DNA was carried out to investigate whether programmed cell death (apoptosis) would be inducible by any of the factors. In UM-SCV-1A cells, interleukin-10 (IL-10) and interleukin-13 (IL-13) caused an approximately 12-fold decrease in DNA synthesis in cells cultured for 72 h (P<0.001), while GM-CSF had no significant effect. TGFbeta showed a significant inhibitory effect on deoxyuridine incorporation (P<0.001), which was 2.0- and 4.2-fold at 48 h and 72 h, respectively. TFG alpha showed a 1.2-fold inhibitory effect on DNA synthesis at 48 h (P<0.01) and a 1.5-fold inhibition at 72 h (P<0.05). Interferon gamma (IFNgamma) showed an inhibitory effect on DNA synthesis (1.3-fold; P<0.01). In UM-SCV-6 cells, both IL-10 and IL-13 showed inhibitory effects on deoxyuridine incorporation (1.3- and 1.4-fold at 48 h, respectively; P<0.001) that were even more pronounced at 72 h (2.4- and 2.5-fold respectively; P<0.001). IFNgamma caused a 3.6-fold inhibition of DNA synthesis by UM-SCV-6 cells at 72 h (P<0.001). Both TFGbeta and TNF alpha inhibited uridine incorporation (3.0- and 1.6-fold at 48 h, respectively; 2.7-fold at 72 h for both factors). GM-CSF inihibited DNA synthesis by UM-SCV-6 cells 1.3- 2.0-fold at 48 h and 72 h, respectively. In dose/response analyses, the effect of INF alpha on DNA synthesis was inhibitory in both cell lines at 48 h, while stimulatory effects were observed at 72 h. Electrophoretic analyses of DNA isolated from cells cultured in the presence or absence of different factors did not reveal DNA fragmentation. All cytokines, with the exception of IFN alpha, showed inhibitory effects on DNA synthesis by vulvar carcinoma cells. Of the factors studied, the recently described interleukins 10 and 13 showed potent inhibition of cell growth, encouraging further investigation on the molecular mechanisms of the observed inhibition. Apoptosis does not seem to be induced in the two vulvar carcinoma cell lines by any of the cytokines studied.
目前对外阴癌的生物学特性和发病机制了解甚少。为了更好地了解这种疾病，我们使用了最近建立的外阴鳞状细胞癌系作为模型。从两名个体分离得到的两个细胞系（UM-SCV-1A和UM-SCV-6）在含10%胎牛血清的培养基中进行体外培养。以放射性标记的尿苷为示踪剂，研究白细胞介素10和13、α干扰素和γ干扰素、粒细胞/巨噬细胞生长刺激因子（GM-CSF）、肿瘤坏死因子α（TNFα）以及转化生长因子β（TGFβ）对细胞增殖的影响。此外，对提取的细胞DNA的分子结构进行研究，以探讨是否有任何一种因子可诱导程序性细胞死亡（凋亡）。在UM-SCV-1A细胞中，白细胞介素10（IL-10）和白细胞介素13（IL-13）使培养72小时的细胞DNA合成减少约12倍（P<0.001），而GM-CSF无显著作用。TGFβ对脱氧尿苷掺入有显著抑制作用（P<0.001），在48小时和72小时分别为2.0倍和4.2倍。TFGα在48小时对DNA合成有1.2倍的抑制作用（P<0.01），在72小时有1.5倍的抑制作用（P<0.05）。γ干扰素（IFNγ）对DNA合成有抑制作用（1.3倍；P<0.01）。在UM-SCV-6细胞中，IL-10和IL-13对脱氧尿苷掺入均有抑制作用（48小时分别为1.3倍和1.4倍；P<0.001），在72小时更为明显（分别为2.4倍和2.5倍；P<0.001）。IFNγ在72小时使UM-SCV-6细胞的DNA合成受到3.6倍的抑制（P<0.001）。TGFβ和TNFα均抑制尿苷掺入（48小时分别为3.0倍和1.6倍；72小时两者均为2.7倍）。GM-CSF在48小时和72小时分别使UM-SCV-6细胞的DNA合成受到1.3至2.0倍的抑制。在剂量/反应分析中，α干扰素在48小时对两种细胞系的DNA合成均有抑制作用，而在72小时观察到刺激作用。对在有或无不同因子情况下培养的细胞分离得到的DNA进行电泳分析，未发现DNA片段化。除α干扰素外，所有细胞因子对外阴癌细胞的DNA合成均有抑制作用。在所研究的因子中，最近报道的白细胞介素10和13对细胞生长有强大的抑制作用，这促使对所观察到的抑制的分子机制进行进一步研究。在所研究的任何一种细胞因子似乎均未诱导这两种外阴癌细胞系发生凋亡。
