十八碳烯酸对人SMMC-7721细胞PI3K-Akt转导信号的影响
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摘要
目的观察十八碳烯酸对人肝癌细胞株SMMC-7721细胞增殖的抑制作用,探讨其对磷酸酰肌醇-3激酶/丝氨酸苏氨酸激酶(PI3K/Akt)转导信号的影响。方法分别给予人SMMC-7721细胞0. 2、0. 4、2. 0 mg/L十八碳烯酸(3个十八碳烯酸组)和10 mg/L 5-氟尿嘧啶(5-FU组)及RPMI1640培养液(对照组),培养48 h;用噻唑蓝(MTT)法检测十八碳烯酸对人SMMC-7721细胞的增殖抑制率;用酶联免疫吸附法(ELISA)检测SMMC-7721细胞中环磷酸腺苷(c AMP)、p53、PI3K、Akt浓度。结果给药后培养48 h检测,十八碳烯酸0. 2、0. 4、2. 0 mg/L组和5-FU组人SMMC-7721细胞增殖抑制率分别升高为25. 88%、41. 32%、65. 90%和39. 17%,均高于对照组,差异有统计学意义(P均<0. 01);十八碳烯酸半数抑制浓度(IC50)为0. 79 mg/L,各给药组c AMP蛋白浓度均高于对照组,p53、PI3K、Akt蛋白浓度均低于对照组(P均<0. 05);且依十八碳烯酸浓度的递升(0. 2 mg/L→0. 4 mg/L→2. 0 mg/L),c AMP蛋白浓度呈递增,p53、PI3K、Akt蛋白浓度呈递减(P均<0. 05)。结论十八碳烯酸具有抑制人SMMC-7721细胞增殖的作用,其机制主要与升高SMMC-7721细胞内c AMP水平,降低p53、PI3K水平,抑制PI3K/Akt信号的活性有关。提示PI3K、p53、Akt或可作为肝癌治疗的潜在靶点。
Objective To study the inhibition effect of the octadecanoic acid on human hepatocellular carcinoma cell line SMMC-7721 cells proliferation and 3-phosphoinositide kinase protein kinase( PI3 K-Akt) transmission signal. Methods The SMMC-7721 cells were respectively treated with 0. 2,0. 4,2. 0 mg/L of octadecanoic acid( three Octadecanoic acid groups) and 10 mg/L of 5-FU( 5-FU group) for 48 hours culture,and the SMMC-7721 cells cultured in RPMI1640 culture medium only were served as control group. Thiazolam blue( MTT) method was used to detect the inhibition effect of the octadecanoic acid on SMMC-7721 proliferation. Enzyme linked immunosorbent assay( ELISA) was used to detect the concentrations of cyclic adenosine monophosphate( c AMP),p53,3-phosphoinositide kinase( PI3 K) and serine/threonineprotein kinase( Akt) in SMMC-7721 cells. Results After 48 hours culture,the proliferation inhibition rates for of octadecanoic acid 0. 2,0. 4,2. 0 mg/L and 5-FU groups( 25. 88%,41. 32%,65. 90% and 39. 17%) were significantly higher than that( 0) for RPMI1640( all P < 0. 01),and the 50% inhibitory concentration( IC50) was 0. 79 mg/L for octadecanoic acid. The concentrations of c AMP protein in each medication group were significantly higher than that in control group,and the concentrations of p53,PI3 K and Akt proteins were significantly lower than that in control group( all P < 0. 05). The concentrations of c AMP protein increased progressively,and the concentrations of p53,PI3 K and Akt proteins decreased progressively with the increase of octadecanoic acid concentrations( 0. 2 mg→0. 4 mg→2. 0 mg/L)( all P < 0. 05). Conclusions Octadecanoic acid has the inhibitory effect on human smmc-7721 cell proliferation,and its mechanism is mainly related to the increase of c AMP level,the decrease of p53 and PI3 K levels in SMMC-7721 cells,and inhibition of activity of PI3 K/Akt signal. It is prompted that p53,PI3 K and Akt maybe served as the potential targets for treatment of liver cancer.
Objective To study the inhibition effect of the octadecanoic acid on human hepatocellular carcinoma cell line SMMC-7721 cells proliferation and 3-phosphoinositide kinase protein kinase( PI3 K-Akt) transmission signal. Methods The SMMC-7721 cells were respectively treated with 0. 2,0. 4,2. 0 mg/L of octadecanoic acid( three Octadecanoic acid groups) and 10 mg/L of 5-FU( 5-FU group) for 48 hours culture,and the SMMC-7721 cells cultured in RPMI1640 culture medium only were served as control group. Thiazolam blue( MTT) method was used to detect the inhibition effect of the octadecanoic acid on SMMC-7721 proliferation. Enzyme linked immunosorbent assay( ELISA) was used to detect the concentrations of cyclic adenosine monophosphate( c AMP),p53,3-phosphoinositide kinase( PI3 K) and serine/threonineprotein kinase( Akt) in SMMC-7721 cells. Results After 48 hours culture,the proliferation inhibition rates for of octadecanoic acid 0. 2,0. 4,2. 0 mg/L and 5-FU groups( 25. 88%,41. 32%,65. 90% and 39. 17%) were significantly higher than that( 0) for RPMI1640( all P < 0. 01),and the 50% inhibitory concentration( IC50) was 0. 79 mg/L for octadecanoic acid. The concentrations of c AMP protein in each medication group were significantly higher than that in control group,and the concentrations of p53,PI3 K and Akt proteins were significantly lower than that in control group( all P < 0. 05). The concentrations of c AMP protein increased progressively,and the concentrations of p53,PI3 K and Akt proteins decreased progressively with the increase of octadecanoic acid concentrations( 0. 2 mg→0. 4 mg→2. 0 mg/L)( all P < 0. 05). Conclusions Octadecanoic acid has the inhibitory effect on human smmc-7721 cell proliferation,and its mechanism is mainly related to the increase of c AMP level,the decrease of p53 and PI3 K levels in SMMC-7721 cells,and inhibition of activity of PI3 K/Akt signal. It is prompted that p53,PI3 K and Akt maybe served as the potential targets for treatment of liver cancer.
引文
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[13] Piao Y,Li Y,Xu Q,et al. Association of MTOR and AKT gene polymorphisms with susceptibility and survival of gastric cancer[J].PLo S One,2015,10(8):e0136447.
[14]费洪荣,陈洪蕾,辛晓明,等. Akt抑制剂perifosine抑制胃癌细胞增殖并诱导凋亡的实验研究[J].中国病理生理杂志,2011,27(6):1084-1089.
[15] Yu R,Yu BX,Chen JF,et al. Anti-tumor effects of Atractylenolide I on bladder cancer cells[J]. J Exp Clin Cancer Res,2016,35:40.
[2]俞发荣,张诗爽,张璟,等.十二碳烯酸对人乳腺癌细胞株BT-20环磷酸腺苷、环磷酸鸟苷水平的影响[J].中国临床研究,2016,29(4):482-484.
[3]刘玉梅,祖桂芳,赵晓红,等. Akt/PKB抗凋亡机制与天然活性物质的抗肿瘤作用[J].生命科学,2011,23(1):1-6.
[4]黄秀兰,崔国辉,周克元. PI3K-Akt信号通路与肿瘤细胞凋亡关系的研究进展[J].癌症,2008,21(3):331-336.
[5]王迎春,王金华.ω-3多不饱和脂肪酸对肿瘤细胞侵袭转移的影响及机制研究进展[J].肿瘤学杂志,2016,22(3):236-241.
[6]童茜茜,徐嘉杰,王峰,等.小球藻精油对人结肠癌细胞增殖和凋亡的作用[J].中国食品学报,2016,16(5):25-31.
[7]梁盈,刘颖,王荣,等.米糠油不饱和脂肪酸对肝癌细胞HepG2克隆及迁移能力的影响[J].现代食品科技,2015,31(11):7-12,50.
[8]薛山. N-3PUFA的抗癌功效及其生物学作用机制[J].中国食品添加剂,2016(7):200-206.
[9]耿丽晶,曲兴源,孙竹萍,等.海洋多不饱和脂肪酸DHA抑制肿瘤机制的研究进展[J].食品工业科技,2013,34(22):385-391.
[10]王莉梅,刘睿杰,金青哲,等.多不饱和脂肪酸在癌症发生中的作用机制研究进展[J].中国油脂,2014,39(8):37-41.
[11]朱文劲,许庆文,鲁珏,等.ω-3多不饱和脂肪酸对大鼠结直肠癌的影响及与脂质代谢相关基因的研究[J].黑龙江医学,2015,39(4):346-349.
[12]范亮亮,马立宁,彭元亮,等. PI3K/AKT信号通路与心力衰竭[J].生命科学研究,2015,19(1):85-90.
[13] Piao Y,Li Y,Xu Q,et al. Association of MTOR and AKT gene polymorphisms with susceptibility and survival of gastric cancer[J].PLo S One,2015,10(8):e0136447.
[14]费洪荣,陈洪蕾,辛晓明,等. Akt抑制剂perifosine抑制胃癌细胞增殖并诱导凋亡的实验研究[J].中国病理生理杂志,2011,27(6):1084-1089.
[15] Yu R,Yu BX,Chen JF,et al. Anti-tumor effects of Atractylenolide I on bladder cancer cells[J]. J Exp Clin Cancer Res,2016,35:40.