摘要
目的:探究半枝莲含药血清对膀胱癌T24细胞凋亡和黏附力的影响。方法:SPF级雄性Wistar大鼠20只,随机分为无药血清组、半枝莲低剂量组、半枝莲中剂量组、半枝莲高剂量组,每组5只,分别以蒸馏水及半枝莲1.5g/(kg·d)、3g/(kg·d)、6g/(kg·d)腹腔注射灌胃,连续3d。末次灌胃后抽取心主动脉血,经灭活处理及微孔过滤或获得半枝莲含药血清。体外培养膀胱癌T24细胞株,分为无药血清组、半枝莲(低、中、高剂量)组,分别对应各组动物血清,以无药血清及含10%的药物血清培养。使用CCK-8法检测细胞增殖情况;流式细胞仪检测各组细胞凋亡率及周期分布;MTT酶标仪法检测各药物浓度下细胞的黏附率;以蛋白印记免疫法检测凋亡相关蛋白[B-细胞淋巴瘤因子2(Bcl-2)抗体、BCL2关联X蛋白(Bax)抗体、胱天蛋白酶3(CASP3)]的表达。结果:与无药血清组比较,半枝莲低剂量组24h及48h的增殖抑制率及凋亡率、细胞G0/G1期、黏附率增加(P<0.05);与半枝莲低剂量组比较,半枝莲中剂量组24h及48h的增殖抑制率及凋亡率、细胞G0/G1期、黏附率增加(P<0.05);与半枝莲中剂量组比较,半枝莲高剂量组24h及48h的增殖抑制率及凋亡率、细胞G0/G1期、黏附率增加(P<0.05)。与无药血清组比较,半枝莲低剂量组Bcl-2相对表达量下降,Bax、CASP3相对表达量增加(P<0.05);与半枝莲低剂量组比较,半枝莲中剂量组Bcl-2相对表达量下降,Bax、CASP3相对表达量增加(P<0.05);与半枝莲中剂量组比较,半枝莲高剂量组Bcl-2相对表达量下降,Bax、CASP3相对表达量增加(P<0.05)。结论:半枝莲含药血清体外培养膀胱癌T24细胞,可有效抑制细胞增殖、降低黏附力,并能诱导细胞凋亡,具有作为抗膀胱癌药物研发的价值。
Objective:To investigate the efeete of drug containing serum of scutellaria barbata on apoplosis and adhesion of bladder cancer T24 cells.Methods:20 male Wistar rats of SPF grade were randomly divided into drug free serum group,scute-laria barbata low-dose group,scuellria barbata medium-dose group and scutellaria barbata high-dose group,5 rats in each group,and were gavaged with dilld water and seutellaria barbata 1.5g/(kg·d),3g/(kg·d)and 6g/(kg·d)intraperito-neally for three consecutive days.Blood from the heart and aorta was extracted afer the last inactivation treatment and micro-porous filtration or the drug containing serum was obtained.Bladder cancer T24 cell lines were cultured in vitro and divided in-to drug-free serum group and scutellaria barbata(low,medium and high dose)groups,corresponding to animal serum of each group,and cultured with drug-free serum and 10%drug-containing serum.Detection of cell proliferation using the CCK-8 meth-od.Flow eytometry to detect the apoplosis rate and eyele distribuion of cells in each group.Dection of ell adhesion rate at each drug concentration by MTT enzyme marker assay.The expression of apoptosis-related proteins[B-cell lymphoma-2(Bel-2)antibody,Recombinant Bel-2 Associated X Protein(Bax)antibody,Caspase 3(CASP3)]was dected by protein-blot immu-noassay.Results:Compared with the drug free serum group,the proliferation inhibition rate at 24 hours and 48 hours,apoplosis rate,G0/G1 phase and adhesion rate were increased in scutellaria barbata low dose group(P<0.05).Compared with scutel-laria barbata low dose group,the proliferation inhibition rate at 24 hours and 48 hours,apoplosis ralte,G0/G1 phase and adhe-sion rate were increased in scutellaria barbata medium dose group(P<0.05).Compared with the scutellaria barbata medium dose group,the proliferation inhibition rate,apoptosis rate,G0/G1 phase and adhesion rate were increased in the scutellaria barbata high dose group at 24 hours and 48 hours(P<0.05).Compared with the drug-free serum group,the relaive expres-sion of Bel-2 was decreased,and the relative expression of Bax and CASP3 was increased in the scutellaria barbata low dose group(P<0.05).Compared with scutellaria barbata low dose group,the relative expression of Bel-2 in scutellaria barbata me-dium dose group was decreased,while the relative expression of Bax and CASP3 was increased(P<0.05).Compared with scutellaria barbata medium dose group,the relative expression of Bc-2 in scutellaria barbata high dose group was deereased,while the relative expression of Bax and CASP3 was increased(P<0.05).Conclusion:The culture of bladder cancer T24 cells in vitro with the drug-containing serum of scutellaria barbata can efectively inhibit cell proliferation,reduce adhesion and induce cell apoptosis,which has the value of developing anti-bladder cancer drugs.
作者
孙业香
王潇潇
王丽华
吕常旭
SUN Yexiang;WANG Xiaoxiao;WANG Lihua(Yantai Nanshan University,Longkou City,Shandong Province 265713)
出处
《医学理论与实践》
2024年第3期371-374,共4页
The Journal of Medical Theory and Practice
基金
2022年烟台南山学院青年基金(NS2022Q12)。
关键词
膀胱癌
半枝莲含药血清
凋亡
细胞黏附力
Bladder cancer
Medicated serum of scutellaria barbata
Apoptosis
Cell adhesion
