Tanaka Kei, Watanabe Momoe, Matsushima Miho, Matsuzawa Yukiko, Izawa Tomoko, Nagashima Takashi, Kobayashi Yoichi, Iwashita Mitsutoshi
Department of Obstetrics and Gynecology, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka, Tokyo 181-8611, Japan.
Department of Obstetrics and Gynecology, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka, Tokyo 181-8611, Japan.
Growth Horm IGF Res. 2018 Aug;41:34-41. doi: 10.1016/j.ghir.2018.07.001. Epub 2018 Jul 4.
Trophoblast survival is regulated by cytokines and growth factors. While the pharmacological levels (10-100 ng/mL) of tumor necrosis factor (TNF)- α affect trophoblasts survival in vitro, the effects of the physiological levels (1-10 pg/mL) of TNF-α remain unknown. We investigated the effects of the physiological levels of TNF-α on proliferation and apoptosis of human trophoblast cells by using BeWo cells. Insulin-like growth factor (IGF)-I is also a potent regulator of trophoblast survival and has been known to exert synergistic effects with other hormones. The interaction of IGF-I and TNF-α on BeWo cells survival was also examined.
After incubating BeWo under the presence of TNF-α (10-10 pg/mL) and IGF-I (10 ng/mL), we assessed cell number by WST-1 assay and cell proliferation by BrdU uptake assay and immunocytochemistry with anti-Ki67 antibody. Apoptosis was evaluated by TUNEL assay and caspase-3, 8 activity assays.
Under the presence of IGF-I, cell number, BrdU uptake, and Ki-67 expression of BeWo were dose-dependently enhanced by low TNF-α (10-10 pg/mL), while no such effects were detected without IGF-I. Higher levels of TNF-α (10-10 pg/mL) showed inhibiting effects on cell number and cell proliferation. The number of TUNEL positive cells were decreased and caspase activities were suppressed by lower levels (10-10 pg/mL) of TNF-α and IGF-I independently. Higher levels of TNF-α (10-10 pg/mL) showed promoting effects on apoptosis irrespective of IGF-I.
The physiological levels of TNF-α and IGF-I had synergetic effects on enhancing cell proliferation and also independently inhibited apoptosis of Bewo cells.
滋养层细胞的存活受细胞因子和生长因子调控。虽然肿瘤坏死因子（TNF）-α的药理水平（10 - 100 ng/mL）会影响体外滋养层细胞的存活，但TNF-α生理水平（1 - 10 pg/mL）的影响尚不清楚。我们使用BeWo细胞研究了TNF-α生理水平对人滋养层细胞增殖和凋亡的影响。胰岛素样生长因子（IGF）-I也是滋养层细胞存活的有效调节剂，并且已知其与其他激素发挥协同作用。还研究了IGF-I和TNF-α对BeWo细胞存活的相互作用。
在TNF-α（10 - 10 pg/mL）和IGF-I（10 ng/mL）存在的情况下孵育BeWo细胞后，我们通过WST-1法评估细胞数量，通过BrdU摄取试验和用抗Ki67抗体的免疫细胞化学评估细胞增殖。通过TUNEL试验和半胱天冬酶-3、8活性测定评估细胞凋亡。
在IGF-I存在的情况下，低剂量TNF-α（10 - 10 pg/mL）可使BeWo细胞数量、BrdU摄取和Ki-67表达呈剂量依赖性增加，而在没有IGF-I的情况下未检测到此类作用。较高水平的TNF-α（10 - 10 pg/mL）对细胞数量和细胞增殖显示出抑制作用。较低水平（10 - 10 pg/mL）的TNF-α和IGF-I可独立降低TUNEL阳性细胞数量并抑制半胱天冬酶活性。无论有无IGF-I，较高水平的TNF-α（10 - 10 pg/mL）均显示出促进细胞凋亡的作用。
TNF-α和IGF-I的生理水平对增强细胞增殖具有协同作用，并且还可独立抑制Bewo细胞的凋亡。
