骆玉霜，格日力，沈存芳，李　燕，范宝华

(1.青海大学高原医学研究中心，青海 西宁 810001；2.青海大学附属医院肿瘤内科，青海 西宁 810001)

低氧下Crocin联合顺铂对人胃癌BGC-823细胞增殖的影响*

骆玉霜1，2，格日力1△，沈存芳2，李燕2，范宝华2

(1.青海大学高原医学研究中心，青海 西宁 810001；2.青海大学附属医院肿瘤内科，青海 西宁 810001)

摘要目的探讨藏红花素联合顺铂(Crocin+DDP)低氧下对人胃癌细胞增殖的影响。方法实验分对照组、C1组、C2组、Z1组、Z2组、C1+Z1组、C1+Z2组、C2+Z1组、C2+Z2组。在低氧环境下培养人胃癌BGC-823细胞，经Crocin、DDP处理后，采用高倍显微镜观察细胞形态变化；各组药物处理胃癌BGC-823细胞后，利用MTT检测细胞增殖情况。结果1)从形态学观察，2%低氧下BGC-823细胞大量死亡，5%低氧下数量减少，MTT示：低氧下48 h BGC-823细胞OD值低于常氧下。2)低氧下DDP处理BGC-823细胞后从形态学观察：细胞固缩、细胞数量减少。MTT示：低氧条件下C1、C2抑制率(分别为30.2%、52.7%)低于在常氧下抑制率(分别为34.9%、59.4%)，P<0.01。3)从形态学观察，常氧下16 mg/mL以下Crocin作用BGC-823细胞后，细胞形态无明显改变，而在低氧下数量增多。MTT示：在低氧下Z2组OD值为0.657±0.038，与低氧对照组(OD值0.554±0.028)比较，存活的细胞增多，P<0.01。4)MTT示：5%低氧下C1+Z1组作用BGC-823细胞48 h，其抑制率为34.7%，与C1组(30.2%)比较，抑制率下降；C1+Z2组抑制率为22.3%，与C1组比较，抑制率下降，P<0.05。C2+Z1组作用BGC-823细胞48 h，抑制率(37.2%)与C2组(52.7%)比较，抑制率下降，P<0.01；C2+Z2组抑制率(51.4%)与C2组比较，抑制率下降。结论1)低氧抑制BGC-823的细胞增殖，低氧可能增加BGC-823细胞对DDP的耐药性。在低氧条件下，一定浓度的Crocin对BGC-823细胞可能起保护作用，抑制DDP对BGC-823细胞抗增殖能力。

关键词藏红花素顺铂细胞增殖低氧BGC-823细胞

项目基金*：青海省科技厅科研(2014-ZJ-737)，青海大学附属医院中青年科研 (ASRF-2014-22)

骆玉霜(1972~)，女，汉族，河南籍，主任医师.△：通迅作者，E-mail：geriligao@hotmail.com

中图分类号R735

文献标识码识码A

DOI：10.13452/j.cnki.jqmc.2015.02.004

AbstractObjectiveTo investigate the effect of Crocin combining with cisplatin on the proliferation of gastric cancer BGC-823 cells under hypoxia.Methods Microscope was used to observe morphological changes of BGC-823 cells after cultured with different concentrations of crocin or cisplatin.Anti-proliferation effect was detected by MTT assay.The trial was set up control group，C1 group(0.8μg/mL DDP)，C2 group(1.6μg/mL DDP)，Z1 group(8mg/mL Crocin)，Z2 group(16 mg/mL Crocin)，C1+Z1 group，C1+Z2 group，C2+Z1 group and C2+Z2 group.Results 1)Morphologically，a large number of BGC-823 cell deaths were observed after 48 h culture in hypoxic(2% O2)condition.A reduction of the amount of cells was observed after 48 h culture in hypoxic(5% O2)condition.The OD value in hypoxia measured by MTT assay was lower than that in normoxia.2)Pyknosis and significant reduction of the amount of BGC-823 cells were observed after being treated by DDP in hypoxia.The inhibitory rates of C1 group and C2 group under hypoxia were 30.2% and 52.7% respectively，which were lower than those under normoxia(34.9%，59.4%)，P<0.01.3)Few alterations of BGC-823 cell morphology were observed after being treated by crocin with concentration below 16 mg/mL under normoxia.The OD value of BGC-823 cells treated by Z2 group was 0.657±0.038，which was higher than that of control group(0.554±0.028)under hypoxia，P<0.01.4)The inhibitory rate of C1+Z1 group was 30.2%，which was lower than that of C1 group；the inhibitory rate of C1+Z2 group was 22.3%，which was lower than that of C1 group(P<0.05)under hypoxia.The inhibitory rate of C2+Z1 group was 50.7%，which was lower than that of C2 group，P<0.01；the inhibitory rate of C2+Z1 group was 51.4%，which was lower than that of C2 group under hypoxia.Conclusions Hypoxia inhibits proliferation of BGC-823 cell and increases drug-resistance to DDP.Crocin protects BGC-823 cells from the ability of DDP on anti-proliferation and anti-apoptosis under hypoxia.

KeywordsCrocinCisplatin(DDP)Cell proliferationHypoxiaBGC-823 cell

收稿日期2015-01-06

THE EFFECT CROCIN COMBINING WITH CISPLATIN

ON PROLIFERATION OF GASTRIC CANCER BGC-823

CELLS UNDER HYPOXIA

Luo Yushuang1，2，Ge Rili1△，Shen Cunfang2，Li Yan2，Fan Baohua2

(1.Research Center for High Altitude Medicine，Qinghai University School of Medicine，Xining，810001；

2.Department of oncology，Affiliated hospital of Qinghai university，Xining 810001，China)

全身化疗是晚期胃癌患者的主要治疗措施。其中顺铂(Cisplnt，DDP)为最常用的化疗药物。到目前为止，由于耐药所致，晚期胃癌的化疗有效率仍然在50%左右。

由于多数中药具有多靶点作用的特点，作为肿瘤多药耐药逆转剂较化学逆转剂有很大的优越性。研究发现，藏红花素(Crocin)不仅可增强低氧耐受，而且对多种肿瘤细胞有抑制作用[1-5]。但Crocin在低氧环境下对肿瘤细胞抑制作用及机制不明。……