Objective:To analyze the effect of spleen on the Treg cells during pregnancy. Methods: The mononuclear cells were separated from the peripheral, spleen and uterus or placenta blood. Flow cytometry was employed to analyze the percent of Treg cells in total T cells in different stages of pregnancy. Immunohistochemical staining was used to make sure the distribution of Treg in spleen in different stages of pregnancy. Results: The results of immunohistochemical staining showed that compared with spleen Treg cells in normal unpregnant mice, spleen Treg cells on day 7 and 14 of pregnancy significantly increased. After splenectomy, peripheral blood and placenta Treg cells on day 7 of pregnancy markedly decreased as compared with the normal pregnancy(P<0.01). And the cells on day 14of pregnancy were markedly recovered as compared with the normal pregnancy. Conclusion: Our study indicated that the spleen and its Treg cells might play important roles in transient tolerance during pregnancy.
目的研究PTEN基因与宫颈癌细胞增殖能力的关系,以探索PTEN基因在宫颈癌发生发展中的可能作用及其机制。方法 Western blot检测PTEN基因在宫颈癌细胞系HeLa、SiHa、C33A和CasKi中的表达。利用脂质体法将人工过表达PTEN载体转染低表达PTEN的宫颈癌细胞系HeLa和C33A,通过MTT方法观察转染后PTEN过表达对宫颈癌细胞增殖的影响。采用多功能过程参数分析仪,检测转染PTEN基因后宫颈癌细胞培养基中乳酸、葡萄糖及谷氨酰胺的含量,观察PTEN基因过表达对宫颈癌细胞代谢的影响。Western blot检测PTEN过表达后宫颈癌细胞中丙酮酸激酶(PKM2)、6-磷酸果糖激酶2(PFKFB3)、谷氨酰胺酶(GLS)、AKT和磷酸化的AKT(pAKT)表达情况。结果与正常宫颈上皮细胞相比,PTEN基因在4种宫颈癌细胞系中低表达。与对照组相比,过表达PTEN的HeLa和C33A细胞克隆中,PTEN基因的水平明显升高。细胞计数和MTT法也显示,过表达PTEN能够明显抑制肿瘤细胞的增殖速率(P<0.05);过表达PTEN基因的细胞培养液中葡萄糖及谷氨酰胺含量显著增多,但乳酸含量减少;同时还发现PTEN抑制了AKT的磷酸化水平并使PKM2、PFKFB3及GLS的表达水平降低。结论 PTEN基因可通过AKT途径调节细胞代谢,从而调节宫颈癌细胞的增殖。PTEN基因有可能成为宫颈癌的诊断和治疗的新靶点。
