Sheng Yu, Liu Hong, Xu Mei-Yu, Jiang Sheng-Hua, Ding Run-Sheng, Lu Wei
Department of Pediatrics, Affiliated Hospital of Nantong University, Nantong 226001, China.
Zhonghua Xue Ye Xue Za Zhi. 2008 May;29(5):296-9.
To investigate the effect of phosphorylated protein kinase C epsilon (pPKC epsilon) on apoptosis of 32D cells induced by sera from patients with aplastic anemia (AA).
The expression of pPKC epsilon and apoptosis in 32D cells were measured by Western blotting and flow cytometry after incubation with sera from healthy individuals (controls, n = 8), patients with severe AA ( SAA, n = 8)and non severe AA (NSAA, n = 6).
After incubation for 0, 12, 24, 36 and 48 hours in the presence of serum and for another 4 hours in medium deprived of serum, the levels of pPKC epsilon in cells in SAA and NSAA group increased gradually, peaked at 24 hours, and then declined (P < 0.05). Compared with that in control group (0.54 +/- 0.08), pPKC epsilon was overexpressed in both SAA group (0.90 +/- 0.10) and NSAA group (0.64 +/- 0.08) (P < 0.05) after 24 hours incubation with serum and subsequent 4 hours without serum. pPKC epsilon level was higher in SAA group than in NSAA group (P < 0.05). A greater proportion of 32D cells showed apoptosis after 24 hours incubation with sera from SAA patients [(4.05 +/- 1.05)%] and subsequent 4 hours incubation without serum than that in controls [(2.45 +/- 0.51)%, P < 0.05], which was correlated with the same serum-induced expression of pPKC epsilon (r = 0.869, P < 0.05). Although the mean level of pPKC epsilon expression was higher in NSAA group than in control group, no significant difference of apoptosis was found between the two groups [(2.45 +/- 0.51)% vs (3.24 +/- 0.56)%, P > 0.05].
Sera from both SAA and NSAA patients could upregulate the expression of pPKC epsilon in 32D cells. The SAA sera induce apoptosis in 32D cells significantly, but the latter do not.
探讨磷酸化蛋白激酶Cε（pPKCε）对再生障碍性贫血（AA）患者血清诱导的32D细胞凋亡的影响。
用健康个体（对照组，n = 8）、重型AA患者（SAA，n = 8）和非重型AA患者（NSAA，n = 6）的血清孵育32D细胞后，通过蛋白质免疫印迹法和流式细胞术检测pPKCε的表达及细胞凋亡情况。
在有血清存在的情况下孵育0、12、24、36和48小时，然后在无血清培养基中再孵育4小时，SAA组和NSAA组细胞中pPKCε水平逐渐升高，在24小时达到峰值，随后下降（P < 0.05）。与对照组（0.54±0.08）相比，血清孵育24小时后再无血清孵育4小时，SAA组（0.90±0.10）和NSAA组（0.64±0.08）中pPKCε均过表达（P < 0.05）。SAA组pPKCε水平高于NSAA组（P < 0.05）。用SAA患者血清孵育24小时后再无血清孵育4小时，32D细胞凋亡比例[（4.05±1.05）%]高于对照组[（2.45±0.51）%，P < 0.05]，这与相同血清诱导的pPKCε表达相关（r = 0.869，P < 0.05）。虽然NSAA组pPKCε表达平均水平高于对照组，但两组凋亡无显著差异[（2.45±0.51）%对（3.24±0.56）%，P > 0.05]。
SAA和NSAA患者的血清均可上调32D细胞中pPKCε的表达。SAA患者血清可显著诱导32D细胞凋亡，而NSAA患者血清则不能。
