青龙衣含药血清抑制胃癌SGC-7901细胞生长和诱导凋亡
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摘要
目的:探讨青龙衣含药血清对胃癌SGC-7901细胞增殖及凋亡的影响,并探讨其作用机制。方法:Wistar大鼠随机分为空白组、青龙衣低剂量(25 g·kg~(-1)·d~(-1)),中剂量(50 g·kg~(-1)·d~(-1)),高剂量(100 g·kg~(-1)·d~(-1))组及顺铂(5 g·kg·d~(-1))组,连续灌胃给药7 d,空白组灌胃给予等体积生理盐水;末次给药2 h后,颈动脉采血,制备青龙衣含药血清;分别采用噻唑蓝(MTT)比色法、流式细胞术、烟酸己可碱(Hoechst)染色、实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测青龙衣含药血清对SGC-7901细胞增殖、凋亡率、凋亡指数、凋亡相关基因及凋亡信号通路蛋白表达的影响。结果:与空白组比较,青龙衣含药血清对SGC-7901细胞的增殖具有明显抑制作用(P<0.05),青龙衣含药血清处理48 h后的早期、晚期凋亡率,凋亡指数及促凋亡基因Bcl-2相关X蛋白(Bax),Bcl-2相关K蛋白(Bak)mRNA表达均升高,抗凋亡基因B淋巴细胞瘤-2(Bcl-2)mRNA表达降低(P<0.05);青龙衣含药血清组β-链蛋白(β-catenin),细胞周期蛋白D1(Cyclin D1)及原癌基因(C-myc)蛋白表达均明显降低(P<0.05)。结论:青龙衣含药血清对胃癌SGC-7901细胞的增殖具有抑制作用,该作用可能与诱导凋亡及抑制分泌型糖蛋白(Wnt)/β-catenin通路的活化有关。
Objective: To explore the effects of serum containing Juglans mandshurica( SCJM) pericarp on proliferation and apoptosis of gastric carcinoma SGC-7901 cells. Method: Wistar rats were randomly divided into blank control group,low( 25 g·kg~(-1)·d~(-1)),medium( 50 g·kg~(-1)·d~(-1)) and high( 100 g·kg~(-1)·d~(-1)) dose J. mandshurica treatment groups and cisplatin group( 5 g·kg~(-1)·d~(-1)). The rats in treatment groups received lavage administration for 7 d,and the rats in blank control group received the equivalent volume of normal saline. 2 h after the last treatment,SCJM was prepared by carotid blood serum; cell proliferation,apoptosis rates,apoptosis index,levels of apoptosis related gene and protein expression were evaluated by methylthiazolyldiphenyl-tetrazolium bromide( MTT) colorimetry,flow cytometry,Hoechst staining,Real-time PCR and Western blot respectively.Result: SCJM could inhibit the proliferation of SGC-7901 cells( P < 0. 05) as compared with blank group,and48 h after treatment by SCJM,early-and late-apoptosis rates,apoptosis index and mRNA levels of Bax and Bak were increased,but mRNA level of B-cell lymphoma-2( Bcl-2) was decreased( P < 0. 05); the protein levels of β-catenin,Cyclin D1 and C-myc in SCJM-treatment groups were lower than those of control group( P < 0. 05).Conclusion: SCJM had inhibitory effect on proliferation of gastric carcinoma SGC-7901 cells,and the effect may be related to the induction of apoptosis and inhibition of Wnt/β-catenin pathway activation.
Objective: To explore the effects of serum containing Juglans mandshurica( SCJM) pericarp on proliferation and apoptosis of gastric carcinoma SGC-7901 cells. Method: Wistar rats were randomly divided into blank control group,low( 25 g·kg~(-1)·d~(-1)),medium( 50 g·kg~(-1)·d~(-1)) and high( 100 g·kg~(-1)·d~(-1)) dose J. mandshurica treatment groups and cisplatin group( 5 g·kg~(-1)·d~(-1)). The rats in treatment groups received lavage administration for 7 d,and the rats in blank control group received the equivalent volume of normal saline. 2 h after the last treatment,SCJM was prepared by carotid blood serum; cell proliferation,apoptosis rates,apoptosis index,levels of apoptosis related gene and protein expression were evaluated by methylthiazolyldiphenyl-tetrazolium bromide( MTT) colorimetry,flow cytometry,Hoechst staining,Real-time PCR and Western blot respectively.Result: SCJM could inhibit the proliferation of SGC-7901 cells( P < 0. 05) as compared with blank group,and48 h after treatment by SCJM,early-and late-apoptosis rates,apoptosis index and mRNA levels of Bax and Bak were increased,but mRNA level of B-cell lymphoma-2( Bcl-2) was decreased( P < 0. 05); the protein levels of β-catenin,Cyclin D1 and C-myc in SCJM-treatment groups were lower than those of control group( P < 0. 05).Conclusion: SCJM had inhibitory effect on proliferation of gastric carcinoma SGC-7901 cells,and the effect may be related to the induction of apoptosis and inhibition of Wnt/β-catenin pathway activation.
引文
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[19]唐化勇,张万生,于航,等.天南星多糖对人肾癌细胞系GRC-1增殖及凋亡作用的影响[J].中国实验方剂学杂志,2016,22(14):155-158.
[20]ZHANG Y S,XIE J Z,ZHONG J L,et al.Acetyl-11-keto-β-boswellic acid(AKBA)inhibits human gastric carcinoma growth through modulation of the Wnt/β-catenin signaling pathway[J].Biochim Biophys Acta,2013,1830(6):3604-3615.
[2]郭锰,魏艳霞,任佳伟,等.青蒿素联合顺铂对胃癌细胞增殖、凋亡及上皮间质转化的影响[J].中成药,2016,38(2):431-434.
[3]JI Q,LUO Y Q,WANG W H,et al.Research advances in traditional Chinese medicine syndromes in cancer patients[J].J Integr Med,2016,14(1):12-21.
[4]LI W,LI C,ZHENG H,et al.Therapeutic targets of traditional Chinese medicine for colorectal cancer[J].J Tradit Chin Med,2016,36(2):243-249.
[5]刘薇,林文翰,季宇彬.青龙衣毒性作用及体外抗肿瘤作用的实验研究[J].中国中药杂志,2004,29(9):887-890.
[6]周媛媛,王栋,牛峰.抗肿瘤中药青龙衣化学成分的研究[J].中草药,2010,41(1):11-14.
[7]胡泽成.青龙衣多糖对结肠癌HCT-116细胞增殖、凋亡及PI3K/Akt信号通路的影响[J].中国实验方剂学杂志,2016,22(18):136-139.
[8]张梅,陈永春.青龙衣醇提物对人白血病K562细胞的基因芯片表达的影响[J].中医药导报,2015,21(18):20-22.
[9]姬艳菊,徐巍.青龙衣提取物对人肝癌细胞株抑制作用的实验研究[J].中医药学报,2014,42(5):30-34.
[10]金凤,林风琴,金敏,等.消癌舒及其含药血清对人宫颈癌Hela细胞体外增殖及迁移的影响[J].中国实验方剂学杂志,2013,19(24):250-254.
[11]蔡宇,陈冰,余绍蕾,等.刺五加提取物含药血清抗肿瘤作用的研究[J].上海中医药杂志,2006,40(1):58-59.
[12]黄臣虎,陆茵,高骁君,等.中药血清药理学研究进展[J].中国实验方剂学杂志,2011,17(10):266-271.
[13]李爱明,赵惠民,揭俊卿.芒柄花黄素对非小细胞肺癌A549细胞增殖及上皮间质转化的抑制作用[J].临床肿瘤学杂志,2015,20(6):481-486.
[14]吴秋芳,唐周舟,张定富.m TOR通路抑制剂依维莫司对宫颈癌Si Ha细胞恶性行为的影响[J].临床肿瘤学杂志,2015,20(8):685-689.
[15]唐鑫,汪俊谷,何新斌.shRNA靶向PAX6对乳腺癌MCF-7细胞增殖、凋亡及上皮间质转化的影响[J].临床肿瘤学杂志,2015,20(11):967-971.
[16]Mc Cubrey J A,Rakus D,Gizak A,et al.Effects of mutations in Wnt/β-catenin,hedgehog,Notch and PI3K pathways on GSK-3 activity-Diverse effects on cell growth,metabolism and cancer[J].Biochim Biophys Acta,2016,1863(12):2942-2976.
[17]SONG L,LI Z Y,LIU W P,et al.Crosstalk between Wnt/β-catenin and Hedgehog/Gli signaling pathways in colon cancer and implications for therapy[J].Cancer Biol Ther,2015,16(1):1-7.
[18]刘映孜,金洁利,黄军,等.粉防己甲素抑制人结肠癌细胞增殖与Wnt/β-catenin信号的关系研究[J].中国药理学通报,2014,30(1):30-34.
[19]唐化勇,张万生,于航,等.天南星多糖对人肾癌细胞系GRC-1增殖及凋亡作用的影响[J].中国实验方剂学杂志,2016,22(14):155-158.
[20]ZHANG Y S,XIE J Z,ZHONG J L,et al.Acetyl-11-keto-β-boswellic acid(AKBA)inhibits human gastric carcinoma growth through modulation of the Wnt/β-catenin signaling pathway[J].Biochim Biophys Acta,2013,1830(6):3604-3615.
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