【作者】 李鹏；

【导师】 何葵；

【作者基本信息】 南华大学 ， 外科学， 2013， 硕士

【摘要】 目的探讨塞来昔布（Celecoxib）抑制肝癌细胞株HepG-2细胞增殖的分子机制。方法提取非同一浓度的Celecoxib (0umol/L、6.25umol/L、12.50umol/L、25.00umol/L、50.00umol/L、100.00umol/L)对肝癌细胞HepG-2进行处理，采用MTT比色法检测肿瘤细胞分别在24h、48h、72h的凋亡情况，观察塞来昔布在体外作用于人肝癌细胞对其细胞增殖的影响。运用Western blot技术，观察HepG-2细胞经塞来昔布处理后对磷脂酰肌醇3-激酶（PI3K）和磷酸化的蛋白激酶-B（p-Akt）及其下游产物Caspase-9、Caspase-3在不同浓度情况下蛋白表达量的变化。结果1.随着Celecoxib作用时间的延长及药物剂量的增加，其抑制HepG-2细胞增殖的作用明显增强，药物剂量为6.25umol/L时24h的抑制率为11.50％。72h的抑制率为30.15％。药物剂量为100.00umol/L时24h的抑制率为46.79％。72h的抑制率为74.28％（P＜0.01），且呈明显剂量依赖及时间依赖性。2.Celecoxib在随着药物浓度增加后，PI3K、p-Akt的蛋白表达量逐渐下降。Celecoxib作用72h后，PI3K在各浓度蛋白相对表达量分别为1.62、1.43、1.09、0.79、0.59。p-Akt在各浓度蛋白相对表达量分别为0.86、0.81、0.72、0.58、0.30（P＜0.05）。而其下游的Caspase-9在Celecoxib作用72h后，各浓度蛋白的相对表达量分别为0.71、0.98、1.21、1.48、1.72。Caspase-3在Celecoxib作用72h后，各浓度蛋白的相对表达量分别为0.80、1.34、1.48、1.74、2.22（P＜0.05）。结论1.Celecoxib能抑制人肝癌细胞HepG-2的增殖。2.Celecoxib可能通过抑制PI3K/Akt信号通路中PI3K、p-Akt蛋白的表达，上调Caspase家族成员的活性，从而抑制人肝癌细胞的增殖。更多还原

【Abstract】 ObjectiveDiscuss celecoxib inhibit the molecular mechanism of liver cancer cell linesHepG-2cell proliferation.MethodsExtracting Celecoxib in different concentration (0umol/L,6.25umol/L,12.50umol/L,25.00umol/L,50.00umol/L, and100.00umol/L) on liver cancer cellHepG-2, determined by MTT colorimetric method to detect the apoptosis of tumorcells respectively in24h,48h,72h and observation of celecoxib in vitro on humanliver cancer cells affect the cell proliferation. Using Western blot technology,observing the HepG-2cell treated with celecoxib on PI3K and p-Akt and theirdownstream products of Caspase-9, Caspase-3under the condition of differentconcentrations of protein expression changes.Results1. Longer duration of Celecoxib effect and the increase of drug dose, the role ofthe HepG-2cell proliferation inhibition enhanced obviously, doses of6.25umol/L for24h, the inhibition rate is11.50%.72h of inhibition rate is30.15%. Doses of100.00umol/L for24h, the inhibition rate are46.79%.72h the inhibition rate is74.28%(P <0.01), and have an obvious dependent on dose and time.2. Celecoxib increased with the drug concentration, PI3K, p-Akt proteinexpression levels gradually decline.After72h Celecoxib’s effect, PI3K proteins invarious concentrations relative expressions are1.62,1.43,1.09,0.79and0.59. P-Aktin the relative expression quantity of protein concentrations is respectively0.86,0.81,0.72,0.81and0.72(p <0.05). And its downstream Caspase-9in Celecoxib’s effect after72h, the relative expression of protein concentration is respectively0.71,0.98,1.21,0.98and1.21. Caspase-3in Celecoxib’s effect after72h, the relative expressionof protein concentration is respectively0.80,1.34,1.48,1.34and1.48(P <0.05).Conclusions1. Celecoxib can inhibit the HepG-2cell proliferation of liver cancer.2. Celecoxib can inhibit the PI3K/Akt signal path in the expression of PI3K,p-Akt protein raise the activity of the Caspase related-members, thus inhibiting humanliver cancer cells’ proliferation.更多还原