拓?fù)涮婵祵Ρ茄拾┮浦擦鰰r辰放射增敏作用的機(jī)制研究
發(fā)布時間：2018-08-09 08:33
【摘要】：目的：通過構(gòu)建人低分化鼻咽癌裸鼠移植瘤模型,從細(xì)胞水平和分子水平兩方面探討拓?fù)涮婵?TPT)時辰放射增敏的作用機(jī)制。 方法：240只BALB/c (nu/nu)裸鼠在統(tǒng)一光照條件下同步化飼養(yǎng),建立統(tǒng)一的生物節(jié)律至少3周。隨后,在裸鼠大腿外側(cè)皮下接種人低分化鼻咽癌細(xì)胞(CNE2),建立裸鼠鼻咽癌移植瘤模型。成瘤并達(dá)到入組標(biāo)準(zhǔn)后,按照隨機(jī)分組的原則,將達(dá)標(biāo)的裸鼠隨機(jī)分為4組：空白對照組、TPT治療組(單化組)、RT組(單放組)、TPT+RT組,每組48只。觀察四個時辰亞組3HAL0(光照后小時,hours after light onset)、9HAL0、15HAL0、21HALO治療后的情況。其中,拓?fù)涮婵到o藥采用單次腹腔注射給藥法,10mg/kg,在各時辰點(diǎn)前30分鐘給入；RT組、TPT+RT組在4個時辰點(diǎn)分別進(jìn)行單次放療,劑量為18Gy；空白對照組在移植瘤達(dá)標(biāo)后不給予任何治療。時辰放療結(jié)束后1小時處死各組的一半裸鼠,剩余裸鼠觀察生長曲線,并測定腫瘤再生長延緩時間(TGD)。按要求獲取腫瘤標(biāo)本后,一部分采用免疫組化法檢測各組腫瘤標(biāo)本中HP-1、γ-H2AX的表達(dá),用免疫組化圖像分析軟件進(jìn)行半定量分析；一部分用DNA瓊脂糖凝膠電泳法檢測拓?fù)洚悩?gòu)酶Ⅰ的表達(dá)；一部分采用流式細(xì)胞技術(shù)檢測細(xì)胞周期DNA含量及凋亡率。本實(shí)驗(yàn)采用完全隨機(jī)單因素方差分析法(ANOVA法)對各組的檢測結(jié)果進(jìn)行統(tǒng)計學(xué)分析；并采用q檢驗(yàn)進(jìn)行組間兩兩比較,檢驗(yàn)其差異性。 結(jié)果：1.各處理因素均對鼻咽癌移植瘤產(chǎn)生不同程度的抑制作用。通過鼻咽癌移植瘤的時辰放射增敏的整體動物試驗(yàn),發(fā)現(xiàn)：相同時辰點(diǎn)治療時,以TPT+RT組對腫瘤的對腫瘤的抑制效果最好。不同時辰亞組治療時,在放射增敏組療效有較明顯差異,15HAL021HAL09HAL03HAL0,以15HAL0整體療效最好。2.免疫組化檢查結(jié)果顯示：在對照組中,HP-1的表達(dá)呈15HAL021HAL09HAL03HAL0,3HAL0與15HAL0時辰亞組比較有統(tǒng)計學(xué)的差異,提示鼻咽癌組織中乏氧狀況具有時間節(jié)律性,腫瘤組織乏氧狀況為3HAL0最明顯,15HAL0較低。同一時辰亞組,TPT+RT組、RT組與對照組之間HP-1及γ-H2AX的表達(dá)差異均有統(tǒng)計學(xué)意義(P0.01)。不同時辰亞組,在TPT+RT組和RT組相同組別中HP-1的表達(dá)：3HAL09HAL021HAL015HAL0,而γ-H2AX的表達(dá)：15HAL021HAL09HAL03HAL0,兩者在15HAL0與3HAL0的表達(dá)水平比較均有顯著性差異(P0.01)。3. DNA瓊脂糖凝膠電泳法檢測DNA拓?fù)洚悩?gòu)酶Ⅰ的表達(dá),呈15HAL021HAL09HAL03HAL0趨勢,15HAL0與3HAL0組間比較有統(tǒng)計學(xué)意義。4.流式細(xì)胞技術(shù)檢測細(xì)胞周期含量及凋亡率,各處理組與對照組比較,S期細(xì)胞比例下降和凋亡指數(shù)增加,與對照組比較有統(tǒng)計學(xué)意義。 結(jié)論：本實(shí)驗(yàn)證實(shí)了拓?fù)涮婵祵Ρ茄拾┮浦擦鼍哂袝r辰放射增敏作用,對腫瘤的時辰放射增敏作用呈現(xiàn)晝夜節(jié)律性,以15HAL0的TPT+RT組對腫瘤的治療效果最好。探討拓?fù)涮婵禃r辰放射增敏的機(jī)制,可能與腫瘤乏氧的時間節(jié)律性,細(xì)胞周期的改變及凋亡,DNA雙鏈的損傷,拓?fù)洚悩?gòu)酶Ⅰ的表達(dá)變化等相關(guān)。
[Abstract]:Objective: to construct a nude mouse model of nude mice with low differentiated nasopharyngeal carcinoma (TPT), and to explore the mechanism of radioactivity sensitization of topotecan (TPT) from the level of cell and molecular level.
Methods: 240 BALB/c (nu/nu) nude mice were raised synchronously under unified light conditions to establish a unified biological rhythm for at least 3 weeks. Then, a human low differentiated nasopharyngeal carcinoma cell (CNE2) was inoculated subcutaneously on the lateral thigh of nude mice, and a nude mice model of nasopharyngeal carcinoma was established. Rats were randomly divided into 4 groups: blank control group, TPT treatment group (single group), group RT (single play group), group TPT+RT, 48 in each group. The four hour subgroup 3HAL0 (hours after light onset) and 9HAL0,15HAL0,21HALO treatment were observed. Among them, topotecan was given a single intraperitoneal injection, 10mg/kg, before 30 hours. In group RT and group TPT+RT, group RT and group TPT+RT were treated with a single radiotherapy at 4 hour points, and the dosage was 18Gy. The blank control group did not give any treatment after the transplant tumor reached the standard. After the end of the radiotherapy, half of the nude mice were killed in each group. The growth curve of the remaining nude mice was observed and the tumor regrowth delay time (TGD) was measured. The tumor markers were obtained according to the requirement. After this, part of the immunohistochemical method was used to detect the expression of HP-1, gamma -H2AX in the tumor specimens of each group. Semi quantitative analysis was carried out by immunohistochemical image analysis software. A part of the expression of topoisomerase I was detected by DNA agarose gel electrophoresis, and a part of the cell cycle DNA content and apoptosis rate were detected by flow cytometry. A complete random single factor analysis of variance (ANOVA) was used to analyze the results of each group, and the difference between the 22 groups was tested by Q test.
Results: 1. all the treatment factors have different inhibitory effects on the transplanted tumor of nasopharyngeal carcinoma. Through the whole animal test of the time radiosensitization of the nasopharyngeal carcinoma, it is found that the TPT+RT group has the best effect on the tumor in the same time point treatment. The results of 15HAL021HAL09HAL03HAL0, the best.2. immunization test with the overall effect of 15HAL0 showed that in the control group, the expression of HP-1 showed a statistically significant difference between the 15HAL021HAL09HAL03HAL0,3HAL0 and the 15HAL0 hour subgroup, suggesting that the hypoxia in the nasopharyngeal carcinoma tissues has a time rhythm and the hypoxic state of the tumor tissue is 3HAL0 The expression of HP-1 and gamma -H2AX between the same time subgroup, the TPT+RT group, the RT group and the control group was statistically significant (P0.01). The expression of HP-1 in the same group of the TPT+RT group and the RT group in the TPT+RT group and the RT group: 3HAL09HAL021HAL015HAL0, and the expression of gamma -H2AX: 15HAL021HAL09HAL03HAL0, both in the TPT+RT and in the RT. The expression level was significantly different (P0.01).3. DNA agarose gel electrophoresis was used to detect the expression of DNA topoisomerase I, showing a trend of 15HAL021HAL09HAL03HAL0. There was a significant difference between 15HAL0 and 3HAL0 group, and.4. flow cytometry was used to detect the cell cycle content and apoptosis rate. Each treatment group was compared with the control group, and the proportion of S phase cells was under the proportion of the control group. The decrease and apoptosis index increased significantly compared with the control group.
Conclusion: this experiment confirmed that topotecan has the time radiosensitizing effect on nasopharyngeal carcinoma transplantation tumor, and has the circadian rhythm of the time radiosensitization of the tumor. The effect of 15HAL0 TPT+RT group on the tumor is the best. Changes and apoptosis, DNA double strand damage, expression of topoisomerase I and so on.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R739.63
[Abstract]:Objective: to construct a nude mouse model of nude mice with low differentiated nasopharyngeal carcinoma (TPT), and to explore the mechanism of radioactivity sensitization of topotecan (TPT) from the level of cell and molecular level.
Methods: 240 BALB/c (nu/nu) nude mice were raised synchronously under unified light conditions to establish a unified biological rhythm for at least 3 weeks. Then, a human low differentiated nasopharyngeal carcinoma cell (CNE2) was inoculated subcutaneously on the lateral thigh of nude mice, and a nude mice model of nasopharyngeal carcinoma was established. Rats were randomly divided into 4 groups: blank control group, TPT treatment group (single group), group RT (single play group), group TPT+RT, 48 in each group. The four hour subgroup 3HAL0 (hours after light onset) and 9HAL0,15HAL0,21HALO treatment were observed. Among them, topotecan was given a single intraperitoneal injection, 10mg/kg, before 30 hours. In group RT and group TPT+RT, group RT and group TPT+RT were treated with a single radiotherapy at 4 hour points, and the dosage was 18Gy. The blank control group did not give any treatment after the transplant tumor reached the standard. After the end of the radiotherapy, half of the nude mice were killed in each group. The growth curve of the remaining nude mice was observed and the tumor regrowth delay time (TGD) was measured. The tumor markers were obtained according to the requirement. After this, part of the immunohistochemical method was used to detect the expression of HP-1, gamma -H2AX in the tumor specimens of each group. Semi quantitative analysis was carried out by immunohistochemical image analysis software. A part of the expression of topoisomerase I was detected by DNA agarose gel electrophoresis, and a part of the cell cycle DNA content and apoptosis rate were detected by flow cytometry. A complete random single factor analysis of variance (ANOVA) was used to analyze the results of each group, and the difference between the 22 groups was tested by Q test.
Results: 1. all the treatment factors have different inhibitory effects on the transplanted tumor of nasopharyngeal carcinoma. Through the whole animal test of the time radiosensitization of the nasopharyngeal carcinoma, it is found that the TPT+RT group has the best effect on the tumor in the same time point treatment. The results of 15HAL021HAL09HAL03HAL0, the best.2. immunization test with the overall effect of 15HAL0 showed that in the control group, the expression of HP-1 showed a statistically significant difference between the 15HAL021HAL09HAL03HAL0,3HAL0 and the 15HAL0 hour subgroup, suggesting that the hypoxia in the nasopharyngeal carcinoma tissues has a time rhythm and the hypoxic state of the tumor tissue is 3HAL0 The expression of HP-1 and gamma -H2AX between the same time subgroup, the TPT+RT group, the RT group and the control group was statistically significant (P0.01). The expression of HP-1 in the same group of the TPT+RT group and the RT group in the TPT+RT group and the RT group: 3HAL09HAL021HAL015HAL0, and the expression of gamma -H2AX: 15HAL021HAL09HAL03HAL0, both in the TPT+RT and in the RT. The expression level was significantly different (P0.01).3. DNA agarose gel electrophoresis was used to detect the expression of DNA topoisomerase I, showing a trend of 15HAL021HAL09HAL03HAL0. There was a significant difference between 15HAL0 and 3HAL0 group, and.4. flow cytometry was used to detect the cell cycle content and apoptosis rate. Each treatment group was compared with the control group, and the proportion of S phase cells was under the proportion of the control group. The decrease and apoptosis index increased significantly compared with the control group.
Conclusion: this experiment confirmed that topotecan has the time radiosensitizing effect on nasopharyngeal carcinoma transplantation tumor, and has the circadian rhythm of the time radiosensitization of the tumor. The effect of 15HAL0 TPT+RT group on the tumor is the best. Changes and apoptosis, DNA double strand damage, expression of topoisomerase I and so on.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R739.63
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 黃秋賢,柳郁;從內(nèi)經(jīng)的時間醫(yī)學(xué)觀察惡性腫瘤的節(jié)律[J];哈爾濱醫(yī)藥;1995年03期
2 李光明;劉延友;朱彬;文世民;郎錦義;王正榮;;鼻咽癌時辰放療與常規(guī)放療的比較[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;2006年03期
3 李穎;朱彬;王躍，
本文編號：2173521
本文鏈接：http://sikaile.net/yixuelunwen/wuguanyixuelunwen/2173521.html
最近更新
教材專著
