本文選題：野生型p誘導(dǎo)的蛋白磷酸酶 + 三氧化二砷��； 參考：《衛(wèi)生研究》2017年03期

【摘要】：目的研究抑制野生型p53誘導(dǎo)的蛋白磷酸酶1(wild-type p53-induced phosphatase 1,WIP1)提高肺腺癌細(xì)胞對(duì)砷劑的敏感性及其分子機(jī)制。方法設(shè)計(jì)并合成WIP1編碼基因的靶向siRNA,以Lipofectimient 2000轉(zhuǎn)染入肺腺癌A549細(xì)胞,從而抑制WIP1蛋白的表達(dá)。運(yùn)用Western-blot檢測(cè)抑制WIP1對(duì)三氧化二砷(As_2O_3)誘導(dǎo)的P53磷酸化蛋白及其下游效應(yīng)因子的影響;采用流式細(xì)胞術(shù)檢測(cè)抑制WIP1后細(xì)胞凋亡情況;以噻唑藍(lán)(MTT)法檢測(cè)抑制WIP1后細(xì)胞的存活率。結(jié)果以siRNA技術(shù)成功將A549細(xì)胞中WIP1 mRNA和蛋白表達(dá)水平均抑制70%左右。相同濃度As_2O_3(5~40μmol/L)處理后,WIP1表達(dá)抑制組細(xì)胞中P53 ser15表達(dá)低于對(duì)照組,而P53 ser46表達(dá)顯著高于對(duì)照組。此外,相同濃度As_2O_3(10~40μmol/L)處理后,WIP1表達(dá)抑制組中P53 ser15下游效應(yīng)因子——P53上調(diào)凋亡誘導(dǎo)蛋白(P53 up-regulated modulator of apoptosis,PUMA)的表達(dá)水平顯著低于對(duì)照組,而P53ser46下游效應(yīng)因子——細(xì)胞周期調(diào)節(jié)蛋白P21的表達(dá)顯著高于對(duì)照組。與之對(duì)應(yīng)的是,相同劑量As_2O_3(5~40μmol/L)作用下,WIP1表達(dá)抑制組細(xì)胞的凋亡率和細(xì)胞死亡率均顯著高于對(duì)照組。結(jié)論抑制肺腺癌細(xì)胞中WIP1的表達(dá)可通過(guò)促進(jìn)P53磷酸化進(jìn)程提高砷引發(fā)肺腺癌細(xì)胞凋亡的效率。
[Abstract]:Objective to study the molecular mechanism of inhibition of wild-type p53 induced protein phosphatase 1 (wild-type p53-induced phosphatase 1 WIP1) on the susceptibility of lung adenocarcinoma cells to arsenic. Methods targeted siRNAs encoding WIP1 gene were designed and synthesized and transfected into lung adenocarcinoma A549 cells with lipofectious 2000 to inhibit the expression of WIP1 protein. Western-blot was used to detect the effect of inhibition of WIP1 on p53 phosphorylated protein and its downstream effector factors induced by arsenic trioxide (as _ S _ 2O _ 3), flow cytometry was used to detect the cell apoptosis after inhibition of WIP1, and MTT method was used to detect the survival rate of cells after inhibition of WIP1. Results the expression of WIP1 mRNA and protein in A549 cells was inhibited by 70% by siRNA technique. The expression of p53 ser15 and p53 ser46 in the control group were significantly lower than those in the control group after treatment with the same concentration of as _ 2O _ 3 (5 ~ 40 渭 mol / L), but the expression of p53 ser46 was significantly higher than that in the control group. In addition, the expression level of p53 up-regulated modulator of apoptosis-inducing protein (Puma) was significantly lower in the group treated with the same concentration of as _ 2O _ 3 (10 ~ 40 渭 mol / L) than in the control group. The expression of P21, the downstream effector factor of P53ser46, was significantly higher than that of the control group. In contrast, the apoptosis rate and cell death rate of the WIP1 expression inhibition group were significantly higher than those of the control group under the same dose of As-Sh _ 2O _ 3 (5 ~ 40 渭 mol / L). Conclusion inhibiting the expression of WIP1 in lung adenocarcinoma cells can increase the efficiency of arsenic induced apoptosis of lung adenocarcinoma cells by promoting p53 phosphorylation.
【作者單位】： 四川農(nóng)業(yè)大學(xué)食品學(xué)院;四川大學(xué)華西公共衛(wèi)生學(xué)院;
【基金】：國(guó)家自然科學(xué)基金面上項(xiàng)目(No.81372945)
【分類號(hào)】：R734.2

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