果王素联合GP化疗对小鼠肺腺癌抑制作用及机制

发布时间：2018-05-04 04:00

本文选题：Lewis肺癌 + 果王素　；参考：《南华大学》2015年硕士论文

【摘要】：目的:观察果王素对Lewis肺癌小鼠的作用以及辅助化疗的作用,并初步探讨其作用机制。方法:40只雄性C57BL/6J小鼠,肺腺癌细胞接种其右后腿后随机分为5组,每组8只小鼠,分别为对照组、果王素组、化疗组、低剂量果王素联合化疗组、高剂量果王素联合化疗组。于肿瘤接种第4天后开始进行药物干预,对照组:予以蒸馏水每天灌胃一次,容积为0.3ml/鼠/次,接种后第5,12,19天各腹腔注射一次,容积为蒸馏水0.4ml/鼠/次;果王素组:予以果王素180mg/kg/次,容积为0.3ml/鼠/次,每天灌胃一次;化疗组:按GP方案,顺铂于接种后第5,12,19天各腹腔注射一次,剂量为3mg/kg/次,容积为0.2ml/鼠/次,吉西他滨于接种后第5,12,19天各腹腔注射一次,剂量为50mg/kg/次,容积为0.2ml/鼠/次;低剂量果王素联合化疗组:化疗方案同化疗组,予以每天灌胃一次,果王素剂量为60mg/kg/次,容积为0.3ml/鼠/次;高剂量果王素联合化疗组:化疗方案同化疗组,予以每天灌胃一次,果王素剂量为180mg/kg/次,容积为0.3ml/鼠/次。肿瘤接种第20天后颈椎脱臼处死全部小鼠。实验过程中观察小鼠移植瘤的生长;统计转移到肺组织结节的数量,计算转移抑制率;测移植瘤的质量、体积,计算抑瘤率;肺组织病理改变用HE染色法观察,免疫组化法测各组移植瘤Survivin、Bcl-2、VEGF的蛋白表达水平,RT-PCR法测定各组移植瘤ERCC1、RRM1、Survivin和Bcl-2 m RNA的表达水平,Western blot检测各组移植瘤ERCC1、RRM1蛋白的表达。结果:1、移植瘤的生长、转移情况各给药组与对照组比较移植瘤质量与体积均有不同程度的降低,高剂量果王素联合化疗组移植瘤质量与体积最小,与对照组相比差异有显著性(P0.01),本研究中对照组、果王素组、化疗组、低剂量果王素联合化疗组、高剂量果王素联合化疗组移植瘤的质量分别为(5.95±0.20)g、(5.06±0.45)g、(3.53±0.52)g、(3.18±0.93)g、(2.29±0.31)g;各组移植瘤的体积分别为(6.66±0.49)cm3、(5.55±0.56)cm3、(3.72±0.39)cm3、(3.41±0.48)cm3、(2.41±0.58)cm3;各组小鼠肺转移结节数分别为(14.6±2.6)、(9.9±1.7)、(9.3±1.8)、(5.7±1.6)、(2.0±1.1),果王素组、化疗组、低剂量果王素联合化疗组、高剂量果王素联合化疗组抑瘤率分别为14.95%、41.77%、44.21%和59.68%,各药物干预组转移的抑制率分别为32.19%、36.3%、60.7%和86.4%。2、药物干预后C57小鼠移植瘤VEGF的表达各用药组的VEGF表达与对照组相比均降低,且高剂量果王素联合化疗组降低最为明显,与对照组比较差异有显著性(P0.01)。各组的VEGF蛋白的表达量分别为(150.97±7.06)、(129.75±6.41)、(111.29±9.89)、(98.84±8.46)、(80.56±6.66)。3、药物干预后小鼠移植瘤凋亡基因Survivin、Bcl-2的表达对照组Survivin、Bcl-2蛋白及m RNA表达高于各药物干预组,各药物干预组中高剂量果王素联合化疗组Survivin、Bcl-2蛋白及m RNA的表达最低,与对照组比差异有统计学意义(P0.01)。各组的Bcl-2 m RNA的表达量分别为(101.59±8.15)、(83.42±5.42)、(68.65±2.47)、(52.93±2.90)、(35.94±5.46);Survivin m RNA的表达量分别为(111.43±9.39)、(92.41±6.98)、(82.02±1.59)、(62.02±3.12)和(53.07±2.67)。各组的Bcl-2蛋白的表达量分别为(148.98±9.61)、(128.92±4.89)、(106.45±7.66)、(97.54±3.55)、(77.89±7.31);Survivin蛋白的表达量分别为(158.88±8.11)、(138.48±9.28)、(125.85±8.92)、(110.44±7.80)、(84.14±11.42)。4、药物干预后小鼠移植瘤耐药基因ERCC1、RRM1的表达各药物干预组RRM1与ERCC1的m RNA及蛋白表达水平均较对照组低,且高剂量果王素联合化疗组RRM1与ERCC1的m RNA及蛋白表达最低,其与对照组比差异有统计学意义(P0.01)。各组ERCC1 m RNA的表达量分别为(104.62±4.50)、(58.59±2.28)、(99.45±4.45)、(82.46±2.57)、(57.68±3.76);各组RRM1 m RNA的表达量分别为(102.30±5.50)、(60.55±3.51)、(98.84±4.44)、(80.66±6.35)、(55.60±3.62)。各组ERCC1蛋白的表达量分别为(86.29±2.84)、(40.04±1.50)、(82.66±2.68)、(43.47±3.42)和(37.03±3.17);各组RRM1蛋白的表达量分别为(97.58±2.46)、(38.36±2.15)、(94.77±3.61)、(57.09±1.68)和(33.29±4.20)。结论:1.果王素具有抑制小鼠Lewis肺癌的生长和转移作用,果王素浓度愈高,抑制效果越好。2果王素能提高肺腺癌细胞对化疗药物的敏感性,其机制可能与降低RRM1与ERCC1的表达有关。
[Abstract]:Objective: To observe the effect of fruit and royal hormone on Lewis lung cancer mice and the role of adjuvant chemotherapy, and to explore its mechanism. Methods: 40 male C57BL/6J mice were randomly divided into 5 groups after inoculation of the right hind leg, 8 mice in each group, which were the control group, the fruit crown group, the chemotherapy group, the low dose of the combined chemotherapy group and the high dose of the chemotherapy group. The combination chemotherapy group. After fourth days of inoculation, the drug intervention began to be carried out in the control group. The control group was treated with distilled water once every day, the volume was 0.3ml/ rats / times, the volume was 0.4ml/ rats / times of distilled water on the 5,12,19 day after inoculation, and the fruit and Royal Group: the fruit Wang Su 180mg/kg/ times, the volume was 0.3ml/ mice / times, gavage every day. Chemotherapy group: chemotherapy group: according to the GP regimen, cisplatin was intraperitoneally injected each time after inoculation on day 5,12,19, the dose was 3mg/kg/ times, the volume was 0.2ml/ rats / times, and gemcitabine was injected once after 5,12,19 day after inoculation, the dose was 50mg/kg/ times, the volume was 0.2ml/ mice / times; the low dose Wang Su combined chemotherapy group: chemotherapy regimen assimilation group, give daily irrigation. Once the stomach, the dose of the fruit and royal hormone was 60mg/kg/ times, the volume was 0.3ml/ rats / times; the high dose of fruit and royal hormone combined chemotherapy group: the chemotherapy regimen was assimilated every day, the dose of the fruit and royal hormone was 180mg/kg/ times, the volume was 0.3ml/ mice / times. All the mice were killed at the dislocation of the cervical vertebra after twentieth days of tumor inoculation. The number of pulmonary nodules, the rate of metastasis inhibition, the mass, volume, and the tumor suppressor rate were measured, the pathological changes of lung tissue were observed by HE staining, and the protein expression levels of Survivin, Bcl-2, VEGF were measured by immunohistochemistry, and the expression of ERCC1, RRM1, Survivin and Bcl-2 m in each group of transplanted tumors was determined by RT-PCR. Level, Western blot was used to detect the expression of ERCC1 and RRM1 protein in each group of transplanted tumors. Results: 1, the growth and metastasis of the transplanted tumor were lower than those of the control group. The mass and volume of the transplanted tumor in the high dose combined chemotherapy group were the smallest, compared with the control group, the difference was significant (P0.01). In the study, the control group, the fruit and royal group, the chemotherapy group, the low dose of the combined chemotherapy group and the high dose of the combined chemotherapy group were (5.95 + 0.20) g, (5.06 + 0.45) g, (3.53 + 0.52) g, (3.18 + 0.93) g and (2.29 + 0.31) g, respectively, and the volume of the graft was (6.66 + 0.49), cm3, cm3, and cm3, respectively. Cm3, (2.41 + 0.58) cm3, the number of pulmonary metastasis nodules of mice in each group was (14.6 + 2.6), (9.9 + 1.7), (9.3 + 1.8), (5.7 + 1.6), (2 + 1.1), fruit and royal group, chemotherapy group, low dose of fruit crown combined chemotherapy group. 2.19%, 36.3%, 60.7% and 86.4%.2, the expression of VEGF expression in the VEGF of C57 mice was lower than that of the control group, and the high dose of the combined chemotherapy group decreased the most significantly (P0.01). The expression of VEGF protein in each group was (150.97 + 7.06), (129.75 + 6.41), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11), (11 1.29 + 9.89), (98.84 + 8.46) and (80.56 + 6.66).3, the apoptosis gene of transplanted tumor in mice was Survivin, the expression of Bcl-2 was Survivin, Bcl-2 protein and m RNA were higher than that of the drug intervention group. The expression of Survivin, Bcl-2 protein and m RNA in the high dose combined chemotherapy group of each drug intervention group was the lowest, and the difference was compared with the control group. Statistical significance (P0.01). The expression of Bcl-2 m RNA in each group was (101.59 + 8.15), (83.42 + 5.42), (68.65 + 2.47), (52.93 + 2.90), (35.94 + 5.46), and Survivin m RNA (111.43 + 9.39), (92.41 + 68.65), (2.47), and (2.47). (128.92 + 4.89), (106.45 + 7.66), (97.54 + 3.55), (77.89 + 7.31), the expression of Survivin protein was (158.88 + 8.11), (138.48 + 9.28), (125.85 + 9.28), (125.85 + 97.54),.4, the drug resistance of mice was treated with drug resistance gene ERCC1, and the expression level of M RNA and protein in RRM1 and ERCC1 in each drug intervention group were both compared. The expression of M RNA and protein in RRM1 and ERCC1 in the combined chemotherapy group was lowest, and the difference was statistically significant (P0.01). The expression of ERCC1 m RNA in each group was (104.62 + 4.50), (58.59 + 2.28), (99.45 + 4.45), (82.46 + 2.57), (57.68 + 3.76), and the expression of RRM1 m RNA was (102.30 + 5.50), respectively. (60.55 + 3.51), (98.84 + 4.44), (80.66 + 6.35), (55.60 + 3.62). The expression of ERCC1 protein in each group is (86.29 + 2.84), (40.04 + 1.50), (82.66 + 2.68) and (6.35), respectively. The higher the growth and metastasis of Lewis lung cancer in mice, the higher the concentration of the fruit, the better the inhibitory effect, the better the.2 fruit hormone can improve the sensitivity of lung adenocarcinoma cell to chemotherapeutic drugs. The mechanism may be related to the reduction of the expression of RRM1 and ERCC1.

【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R734.2

【参考文献】