摘要
目的观察抑制磷脂酰肌醇3-激酶(PI3K)信号通路是否有助于小鼠胚胎干细胞向肠上皮细胞分化。方法用PI3K信号通道抑制剂LY 294002作用小鼠胚胎干细胞,以Western blot法检测干细胞PI3K下游分子GSK3β及其活性磷酸化分子GSK3βs9的表达;以定量PCR检测小鼠胚胎干细胞分化第5日细胞各胚层标志基因的表达和分化第11日细胞移植瘤肠上皮细胞标志基因的表达,并与不加LY 294002对照组比较。结果 Western blot法检测LY 294002组和对照组GSK3βS9/GSK3β灰度比值为0.018±0.005和0.818±0.190,差异有统计学意义(P<0.05);LY294002组分化第5日细胞Cxcr4、Sox17、Sox9、TBX6 mRNA相对表达量分别为12.318±2.216、22.154±4.166、9.466±1.238、2.351±0.314,高于对照组(6.142±1.012、6.232±1.307、3.988±0.978、0.823±0.143),两组比较差异有统计学意义(P<0.05);LY294002组移植瘤Villin、FABP2、Lyz、ChA、Lgr5的mR-NA相对表达量为4.422±1.233、4.362±0.897、4.405±1.004、4.877±0.727、7.318±1.565,与对照组比较差异有统计学意义(P<0.05)。结论抑制PI3K信号通路能促进小鼠胚胎干细胞向肠上皮细胞分化。
Objective To observe whether inhibiting phosphoinositide 3-kinase （PI3K） signaling of mouse embryonic stem cells could promote mouse embryonic stem cells to differentiate into intestinal epithelial cells. Methods Western blot was used to detect the inhibition effect of LY 294002 on PI3K signaling of mouse embryonic stem ceils. Quantitative real-time polymerase chain reaction was used to detect the expression of markers of germ layers in 5-day population derived from mouse embryonic stem cells and the expression of markers for intestinal epithelial cells in the grafts from 11-day population derived from mouse embryonic stem cells. Results There was significant difference between the integrated intensities ratio of GSK313S9/GSK3β in LY 294002 group （0. 018 ±0. 005） and control group （0. 818 ±0. 190） （P 〈0. 05）. The mRNA rative ex-pression levels of Cxcr4, Sox17, Sox9, TBX6 in 5-day population in LY 294002 group were 12. 318 ±2. 216, 22. 154 ±4. 166, 9. 466 ± 1. 238, 2. 351 ±0. 314, higher than those in control group （6. 142 ± 1. 012, 6. 232 ± 1. 307, 3. 988 ±0. 978, 0. 823 ±0. 143） （P 〈0. 05）. Compared with control group, the mRNA expression of Villin, FABP2, Lyz, ChA, Lgr5 in the grafts from LY 294002 group were 4. 422 ± 1. 233, 4. 362 ± 0. 897, 4. 405 ± 1. 004, 4. 877 ± 0. 727, 7. 318 ± 1. 565. There was significant difference between LY 294002 group and control group. Conclusions Inhibiting PI3K signaling of mouse embryonic stem cells could promote mouse embryonic stem cells to differentiate into intestinal epithelial cells.
出处
《新医学》
2013年第6期405-409,共5页
Journal of New Medicine
基金
国家自然基金(No.81000152)
广东省自然资金项目(No.S2012040007724)
广州中医药大学科研创新基金资助(NO.11CX068)
中山大学青年教师培育计划基金资助项目(No.09ykpy10)
关键词
胚胎干细胞
肠上皮细胞
PI3K信号
Embryonic stem cells
Intestinal epithelial cells
Phosphoinositide 3-kinase signaling
