發(fā)布時(shí)間：2019-04-23 08:30

【摘要】：目的探討五氯聯(lián)苯PCB126對(duì)人肝癌細(xì)胞有氧糖酵解的影響和機(jī)制。方法取人肝癌細(xì)胞SMMC-7721,用濃度分別為10~(-11)、10~(-10)、10~(-9)、10~(-8)、10~(-7) mol/L PCB126分別處理24、48 h,并以0.1%DMSO作為對(duì)照組。用試劑盒檢測(cè)培養(yǎng)基中葡萄糖含量和乳酸生成量,以實(shí)時(shí)熒光定量PCR法檢測(cè)丙酮酸激酶M2(PKM2)和有氧糖酵解通路中關(guān)鍵因子葡萄糖轉(zhuǎn)運(yùn)蛋白1(GLUT1)、乳酸脫氫酶(LDHA)、丙酮酸脫氫酶激酶(PDK)的表達(dá)。用RNA干擾技術(shù)敲減PKM2,檢測(cè)其對(duì)培養(yǎng)基中葡萄糖含量、乳酸生成量和有氧糖酵解關(guān)鍵因子表達(dá)的影響。結(jié)果與對(duì)照組相比,人肝癌細(xì)胞SMMC-7721經(jīng)10~(-10)、10~(-9)、10~(-8)mol/L PCB126處理48 h后,培養(yǎng)基中葡萄糖含量明顯下降,乳酸生成量明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05);與對(duì)照組相比,處理48 h后對(duì)細(xì)胞存活率無(wú)明顯影響,72 h時(shí)細(xì)胞存活率明顯升高。PCB126暴露可明顯升高PKM2、GLUT1、LDHA和PDK mRNA水平,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。用PKM2 shRNA敲減PKM2后,與10~(-9) mol/L PCB126處理組相比,培養(yǎng)基中葡萄糖含量升高,乳酸生成量下降,GLUT1、LDHA和PDK mRNA水平明顯降低,差異均有統(tǒng)計(jì)學(xué)意義(P0.01)。結(jié)論 PCB126可通過(guò)PKM2上調(diào)肝癌細(xì)胞GLUT1、LDHA和PDK的表達(dá),從而促進(jìn)有氧糖酵解,這可能與PCB126促進(jìn)肝癌的發(fā)展有關(guān)。
[Abstract]:Objective to investigate the effect and mechanism of pentachlorobiphenyl (PCB126) on aerobic glycolysis of human hepatoma cells. Methods SMMC-7721, cells were treated with 10 ~ (- 11), 10 ~ (- 10), 10 ~ (- 9), 10 ~ (- 8), 10 ~ (- 7) mol/L PCB126 for 24 h and 48 h, respectively. 0.1%DMSO was used as control group. Glucose content and lactic acid production in culture medium were detected by kit, pyruvate kinase M2 (PKM2) and key factors in aerobic glycolysis pathway, glucose transporter 1 (GLUT1) and lactate dehydrogenase (LDHA), were detected by real-time fluorescence quantitative PCR. Expression of pyruvate dehydrogenase kinase (PDK). The effects of PKM2, on glucose content, lactic acid production and the expression of key factors in aerobic glycolysis were detected by RNA interference technique. Results compared with the control group, SMMC-7721 cells were treated with 10 ~ (- 10), 10 ~ (- 9), 10 ~ (- 8) mol/L PCB126 for 48 h. The difference was statistically significant (P0.05); Compared with the control group, 48 h treatment had no significant effect on cell survival rate, and 72 h cell survival rate significantly increased. PCB126 exposure significantly increased the levels of PKM2,GLUT1,LDHA and PDK mRNA, the difference was statistically significant (P0.05). Compared with the 10 ~ (- 9) mol/L PCB126 group, the glucose content in the culture medium increased, the lactic acid production decreased, and the levels of GLUT1,LDHA and PDK mRNA decreased significantly after PKM2 shRNA knockout of PKM2 (P0.01). Conclusion PCB126 can up-regulate the expression of GLUT1,LDHA and PDK through PKM2 and promote aerobic glycolysis, which may be related to PCB126 promoting the development of HCC.
【作者單位】： 山西大學(xué)生物技術(shù)研究所化學(xué)生物學(xué)與分子工程教育部重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(21207084);國(guó)家自然科學(xué)基金(31271516) 山西省自然科學(xué)基金(2014011027-5) 高等學(xué)�？萍紕�(chuàng)新項(xiàng)目(2016122)
【分類號(hào)】：R994.6
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本文編號(hào)：2463298