5-氨基乙酰丙酸光动力疗法调控P53/Caspase9通路抑制乳腺癌荷瘤裸鼠肿瘤生长的作用观察  

Effect of 5-aminolevulinic acid photodynamic therapy on P53/Caspase9 pathway inhibiting tumor growth in breast cancer-bearing nude mice

作　　者：罗红兰[1] 刘凯[2] 朱晓东 姬生威[1] 冯兴勇 刘静[1] LUO Hong-lan;LIU Kai;ZHU Xiao-dong(Department of Oncology,Huanggang Central Hospital,Hubei Province, Huanggang Hubei 438000,China;Imaging Department,Huanggang Central Hospital,Hubei Province,Huanggang Hubei 438000,China)

机构地区：[1]湖北省黄冈市中心医院肿瘤内科,湖北黄冈438000 [2]湖北省黄冈市中心医院影像科,湖北黄冈438000

出　　处：《临床和实验医学杂志》2019年第10期1012-1016,共5页Journal of Clinical and Experimental Medicine

摘　　要：目的观察5-氨基乙酰丙酸(5-ALA)光动力疗法调控肿瘤抑制因子(P53)/半胱氨酸蛋白酶9(Caspase9)通路抑制乳腺癌荷瘤裸鼠肿瘤生长的作用。方法取40只裸小鼠,背部接种乳腺癌细胞株MCF-7制作荷瘤鼠模型,建模成功后随机分为5组,模型组(A组)、单纯激光照射组(B组)、低剂量5-ALA+激光照射组(C组)、中剂量5-ALA+激光照射组(D组)、高剂量5-ALA+激光照射组(E组)。C组、D组、E组分别给予瘤内注射10 mg/kg、20 mg/kg、40 mg/kg的5-ALA生理盐水溶液,A组、B组给予瘤内注射无菌生理盐水溶液,2 h后,B组、C组、D组、E组应用半导体激光治疗仪局部照射瘤体治疗。对比各组干预前、干预14 d时肿瘤体积,并比较B组、C组、D组、E组抑瘤率;苏木精-伊红染色法(HE)观察各组肿瘤组织病理学变化;采用原位末端脱氧核苷酸转移酶介导的d UTP缺口末端标记(TUNEL)检测并对比各组肿瘤细胞凋亡率,分别采用实时荧光定量-聚合酶链反应法和蛋白免疫印迹法检测并对比肿瘤组织中P53、Caspase9 mRNA和蛋白表达情况。结果干预前各组肿瘤体积比较,差异均无统计学意义(P>0. 05),干预14 d各组肿瘤体积与干预前比较均显著较高(P <0. 05),干预14 d后肿瘤体积从小至大依次如下,E组、D组、C组、B组、A组,每两组间比较差异均有统计学意义(P <0. 05);抑瘤率从高至低依次如下,E组、D组、C组、B组,每两组间比较差异均有统计学意义(P <0. 05);组织病理学观察发现,A组瘤细胞丰富、排列拥挤呈条索状、血供丰富,细胞大小不一、核大而深染伴有核分裂相,C～E组肿瘤细胞数量较A组明显减少,细胞排列紊乱,且细胞核固缩、染色质淡染,其中E组变化最为明显,B组变化不明显;肿瘤细胞凋亡率、P53、Caspase9 mRNA和蛋白相对表达量从高至低依次如下,E组、D组、C组、B组、A组,每两组间比较差异均有统计学意义(P <0. 05)。结论 5-ALA光动力疗法可显著Objective To observe the effect of 5-aminolevulinic acid(5-ALA)photodynamic therapy on the tumor growth inhibition in nude mice bearing tumor suppressor(P53)/cysteine protease 9(Caspase9)pathway.Methods 40 nude mice were taken and the breast cancer cell line MCF-7 was inoculated into the tumor-bearing mouse model.After successful modeling,they were randomly divided into 5 groups:model group(group A),laser irradiation group(group B),low-dose 5-ALA+laser irradiation group(group C),medium dose 5-ALA+laser irradiation group(group D),high dose 5-ALA+laser irradiation group(group E).Groups C,D,and E were given intratumoral injection of different doses of 5-ALA physiological saline solution,and group A and B were given intratumoral injection of sterile physiological saline solution,after 2 h,groups B,C,D,and E were treated with a semiconductor laser treatment device for local irradiation.The tumor volume before intervention and 14 days after intervention of each group was compared.The tumor inhibition rates of groups B,C,D,and E were compared.Histopathological changes of tumors in each group were observed by hematoxylin-Eosin staining(HE).The apoptosis rate of tumor cells in each group was detected and compared by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).The expressions of P53 and Caspase9 mRNA and protein in tumor tissues were detected and compared by real-time fluorescence quantitative polymerase chain reaction and Western blotting.Results There was no significant difference in tumor volume between the groups before intervention(P>0.05).The tumor volume after 14 days intervention of each group was significantly higher than that before intervention(P<0.05).The tumor volume after 14 days intervention was as follows:E group,D group,C group,B group,A group,the difference was statistically significant between each 2 groups(P<0.05).Histopathological observation showed that the tumor cells in group A were abundant,arranged in a crowded strip,and the blood supply was abundant.The ce

关 键 词：乳腺癌荷瘤裸鼠 5-氨基乙酰丙酸 肿瘤抑制因子P53 半胱氨酸蛋白酶9 

分 类 号：R737.9[医药卫生—肿瘤]