本文選題：Lewis肺癌 + 果王素��； 參考：《南華大學(xué)》2015年碩士論文

【摘要】：目的:觀察果王素對Lewis肺癌小鼠的作用以及輔助化療的作用,并初步探討其作用機制。方法:40只雄性C57BL/6J小鼠,肺腺癌細胞接種其右后腿后隨機分為5組,每組8只小鼠,分別為對照組、果王素組、化療組、低劑量果王素聯(lián)合化療組、高劑量果王素聯(lián)合化療組。于腫瘤接種第4天后開始進行藥物干預(yù),對照組:予以蒸餾水每天灌胃一次,容積為0.3ml/鼠/次,接種后第5,12,19天各腹腔注射一次,容積為蒸餾水0.4ml/鼠/次;果王素組:予以果王素180mg/kg/次,容積為0.3ml/鼠/次,每天灌胃一次;化療組:按GP方案,順鉑于接種后第5,12,19天各腹腔注射一次,劑量為3mg/kg/次,容積為0.2ml/鼠/次,吉西他濱于接種后第5,12,19天各腹腔注射一次,劑量為50mg/kg/次,容積為0.2ml/鼠/次;低劑量果王素聯(lián)合化療組:化療方案同化療組,予以每天灌胃一次,果王素劑量為60mg/kg/次,容積為0.3ml/鼠/次;高劑量果王素聯(lián)合化療組:化療方案同化療組,予以每天灌胃一次,果王素劑量為180mg/kg/次,容積為0.3ml/鼠/次。腫瘤接種第20天后頸椎脫臼處死全部小鼠。實驗過程中觀察小鼠移植瘤的生長;統(tǒng)計轉(zhuǎn)移到肺組織結(jié)節(jié)的數(shù)量,計算轉(zhuǎn)移抑制率;測移植瘤的質(zhì)量、體積,計算抑瘤率;肺組織病理改變用HE染色法觀察,免疫組化法測各組移植瘤Survivin、Bcl-2、VEGF的蛋白表達水平,RT-PCR法測定各組移植瘤ERCC1、RRM1、Survivin和Bcl-2 m RNA的表達水平,Western blot檢測各組移植瘤ERCC1、RRM1蛋白的表達。結(jié)果:1、移植瘤的生長、轉(zhuǎn)移情況各給藥組與對照組比較移植瘤質(zhì)量與體積均有不同程度的降低,高劑量果王素聯(lián)合化療組移植瘤質(zhì)量與體積最小,與對照組相比差異有顯著性(P0.01),本研究中對照組、果王素組、化療組、低劑量果王素聯(lián)合化療組、高劑量果王素聯(lián)合化療組移植瘤的質(zhì)量分別為(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;各組移植瘤的體積分別為(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;各組小鼠肺轉(zhuǎn)移結(jié)節(jié)數(shù)分別為(14.6±2.6)、(9.9±1.7)、(9.3±1.8)、(5.7±1.6)、(2.0±1.1),果王素組、化療組、低劑量果王素聯(lián)合化療組、高劑量果王素聯(lián)合化療組抑瘤率分別為14.95%、41.77%、44.21%和59.68%,各藥物干預(yù)組轉(zhuǎn)移的抑制率分別為32.19%、36.3%、60.7%和86.4%。2、藥物干預(yù)后C57小鼠移植瘤VEGF的表達各用藥組的VEGF表達與對照組相比均降低,且高劑量果王素聯(lián)合化療組降低最為明顯,與對照組比較差異有顯著性(P0.01)。各組的VEGF蛋白的表達量分別為(150.97±7.06)、(129.75±6.41)、(111.29±9.89)、(98.84±8.46)、(80.56±6.66)。3、藥物干預(yù)后小鼠移植瘤凋亡基因Survivin、Bcl-2的表達對照組Survivin、Bcl-2蛋白及m RNA表達高于各藥物干預(yù)組,各藥物干預(yù)組中高劑量果王素聯(lián)合化療組Survivin、Bcl-2蛋白及m RNA的表達最低,與對照組比差異有統(tǒng)計學(xué)意義(P0.01)。各組的Bcl-2 m RNA的表達量分別為(101.59±8.15)、(83.42±5.42)、(68.65±2.47)、(52.93±2.90)、(35.94±5.46);Survivin m RNA的表達量分別為(111.43±9.39)、(92.41±6.98)、(82.02±1.59)、(62.02±3.12)和(53.07±2.67)。各組的Bcl-2蛋白的表達量分別為(148.98±9.61)、(128.92±4.89)、(106.45±7.66)、(97.54±3.55)、(77.89±7.31);Survivin蛋白的表達量分別為(158.88±8.11)、(138.48±9.28)、(125.85±8.92)、(110.44±7.80)、(84.14±11.42)。4、藥物干預(yù)后小鼠移植瘤耐藥基因ERCC1、RRM1的表達各藥物干預(yù)組RRM1與ERCC1的m RNA及蛋白表達水平均較對照組低,且高劑量果王素聯(lián)合化療組RRM1與ERCC1的m RNA及蛋白表達最低,其與對照組比差異有統(tǒng)計學(xué)意義(P0.01)。各組ERCC1 m RNA的表達量分別為(104.62±4.50)、(58.59±2.28)、(99.45±4.45)、(82.46±2.57)、(57.68±3.76);各組RRM1 m RNA的表達量分別為(102.30±5.50)、(60.55±3.51)、(98.84±4.44)、(80.66±6.35)、(55.60±3.62)。各組ERCC1蛋白的表達量分別為(86.29±2.84)、(40.04±1.50)、(82.66±2.68)、(43.47±3.42)和(37.03±3.17);各組RRM1蛋白的表達量分別為(97.58±2.46)、(38.36±2.15)、(94.77±3.61)、(57.09±1.68)和(33.29±4.20)。結(jié)論:1.果王素具有抑制小鼠Lewis肺癌的生長和轉(zhuǎn)移作用,果王素濃度愈高,抑制效果越好。2果王素能提高肺腺癌細胞對化療藥物的敏感性,其機制可能與降低RRM1與ERCC1的表達有關(guān)。
[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.

【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R734.2

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