董学易①, 古 强, 孙 涛, 赵 楠, 赵秀兰, 倪春生, 车 娜, 孙保存②. 转化生长因子-β和干扰素-γ对黑色素瘤细胞增殖迁移和侵袭的影响作用研究*[J]. 中国肿瘤临床, 2010, 37(3): 134-137. DOI: 10.3969/j.issn.1000-8179.2010.03.004
引用本文: 董学易①, 古 强, 孙 涛, 赵 楠, 赵秀兰, 倪春生, 车 娜, 孙保存②. 转化生长因子-β和干扰素-γ对黑色素瘤细胞增殖迁移和侵袭的影响作用研究*[J]. 中国肿瘤临床, 2010, 37(3): 134-137. DOI: 10.3969/j.issn.1000-8179.2010.03.004
DONG Xueyi, GU Qiang, SUN Tao, ZHAO Nan, ZHAO Xiulan, NI Chunsheng, CHE Na, SUN Baocun. Effects of TGF-β and IFN-γ on the Proliferation, Migration and Invasion of Melanoma Cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(3): 134-137. DOI: 10.3969/j.issn.1000-8179.2010.03.004
Citation: DONG Xueyi, GU Qiang, SUN Tao, ZHAO Nan, ZHAO Xiulan, NI Chunsheng, CHE Na, SUN Baocun. Effects of TGF-β and IFN-γ on the Proliferation, Migration and Invasion of Melanoma Cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(3): 134-137. DOI: 10.3969/j.issn.1000-8179.2010.03.004

转化生长因子-β和干扰素-γ对黑色素瘤细胞增殖迁移和侵袭的影响作用研究*

Effects of TGF-β and IFN-γ on the Proliferation, Migration and Invasion of Melanoma Cells

  • 摘要: 目的:观察转化生长因子-β(TGF-β)和干扰素-γ(IFN-γ)对黑色素瘤细胞的增殖、迁移和侵袭能力的影响。方法:体外培养黑色素瘤细胞,待细胞长到80% 融合度时,加入TGF-β(浓度为5ng/mL)和IFN-γ(浓度为10ng/mL)因子,将黑色素瘤细胞分为对照组、TGF-β组、IFN-γ组和TGF-β+IFN-γ组,然后各组分别采用磺酰罗丹明B(SRB )增殖测定法观察黑色素瘤细胞培养后8h、16h、24h、32h、40h 和48h 时间点的增殖情况,于540nm吸光度下比色读取吸光度(A540)来表示细胞的增殖变化。采用200 μ L 吸头对各组细胞的表面进行划痕处理,随后于12h 观察划痕之间距离(即代表肿瘤细胞迁移的能力)的变化。采用测Transwell 系统孔外膜上细胞的595nm吸光度(A595)来表示肿瘤细胞的侵袭能力。采用明胶酶谱法观察各组肿瘤细胞的MMP-2、MMP-9 活性水平,用图像分析软件测定聚丙烯酰胺凝胶中MMP-2、MMP-9 的条带灰度(代表MMPs的活性水平)。 结果:TGF-β 可以增强黑色素瘤细胞的增殖、迁移和侵袭能力(P<0.05),IFN-γ 则抑制黑色素瘤细胞的增殖、迁移和侵袭能力(P<0.05),二者同时处理则对黑色素瘤细胞的影响作用微弱或消失(P>0.05)。 结论:体外实验中,TGF-β 可以干扰IFN-γ 对黑色素瘤细胞的增殖、迁移和侵袭能力的抑制作用,本研究可为炎症和肿瘤的相互关系研究奠定实验基础。

     

    Abstract: Objective: To investigate the influence of TGF-β and IFN-γ on the prol i feration, migration and invasion of melanoma cells. Methods:Melanoma cells were cultured in vitro. When tumor cells were confluent about 80% degree, cytokines were added into cel l cul ture media. The concentration of TGF-β and IFN-γ was 5ng/mL and 10ng/mL, respectively. Melanoma cel ls were divided into free-cytokine group, TGF-β group, IFN-γ group, TGF-β and IFN-γ group. Tumor cel ls in each group were then incubated for 8h, 16h, 24h, 32h, 40h and 48h, respectively. After incubation, fixing and staining with SRB, the optical densities and percentage viability were then determined by absorption at 540 nm (A 540 ). The scarification of tumor cells in each group on the surface was created by a 200 μ L pipette tube. The moti l i ty of tumor cel ls in each group was assessed by measuring the distance between scarifications. The speed of the scuffing closure was monitored after 12h. The invasive ability of melanoma cells was observed by transwell cultivation. The tumor cells that invaded through the Matrigel and adhered to the bottom of the outside membrane were determined by absorption at 595 nm (A 595 ). Gelatin zymography assay was used to examine the levels of matrix metalloproteinases- 2 (MMP- 2) and matrix metalloproteinases- 9 (MMP- 9) activity when the tumor cells were treated with cytokines after24h. MMP- 2 and MMP- 9 activity was demonstrated by gradation in the sodium dodecyl sulfate polyacryl -amide gel electrophoresis (SDS-PAGE) gelatin. MMP-2 and MMP- 9 activity was determined by Image analysis Software. Results: TGF-β promoted the prol i feration, migration and invasion of melanoma cel ls (P<0.05). However, IFN-γ inhibi ted the prol i feration, migration and invasion of melanoma cel ls (P<0.05). The effect weakened or disappeared when both of them were used (P>0.05). Conclusion:In vi tro, TGF-β may affect the inhibi tory effect of IFN-γ on the prol i feration, migration and invasion of melanoma cel ls. This study provided a better understanding of the relationship between tumor and inflammatory factors and established a good ba-sis for future research.

     

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