超声检测肝癌血管分级与血管生成的关系及射频消融术对兔肝VX2残余肿瘤的研究

发布时间：2018-05-11 22:36

本文选题：肝细胞癌 + 微血管密度　；参考：《大连医科大学》2014年硕士论文

【摘要】：目的:超声观察肝癌血流分级及其血流情况，并探讨缺氧诱导因子-1α(hypoxiainducible factor-1alpha，HIF-1α)与微血管密度(microvessel density，MVD)在肝癌组织中的表达及二者间关系。方法:对60例经病理证实的原发性肝癌手术切除术前进行二维及彩色多普勒超声检查，进行血流分级，通过免疫组织化学的方法检测HIF-1α、MVD在肝癌组织中的表达。结果:肝癌超声血流分级为I级时HIF-1a、MVD表达率分别为（64.37±3.02）、（39.32±3.71），血流分级为II级时HIF-1a、MVD表达率分别为（79.94±2.97）、（62.25±3.80），血流分级为III级时HIF-1a、MVD表达率分别为（92.61±3.10）、（85.16±2.33），超声肝癌血流等级和HIF-1α、MVD相关，在肝癌组织中当血流分级为III级时HIF-1a、MVD表达率最高，分别为92.61%±3.10、85.16%±2.33，在正常肝组织中HIF-1a、MVD表达率分别为12.28%±7.21、11.33%±7.21，二者比较具有显著性差异(P0.05)。结论: HIF-1a的表达与肝癌新生血管的生成密切相关，MVD高表达、HIF-1α表达越高，彩色多普勒显示血流越丰富，肝癌超声血流分级能够为临床诊治肝癌、评估疗效提供理论依据。目的:研究兔肝VX2瘤射频消融治疗（Radiofrequency Ablation，RFA）后残癌中胰岛素生长因子-II（Insulin-like growth factor,IGF-II）、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达。方法:实验兔28只，3只VX2荷瘤兔。2只作为传代兔，余23只实验兔行超声引导下肝内瘤块植入，建立VX2肝癌模型。从荷瘤兔切取肿瘤周边生长旺盛的鱼肉样组织，随机取2只实验兔，于双腿内侧局麻后，手术刀破皮，用镊子将瘤块送入皮下，，传代（传代兔）。超声引导下将VX2瘤粒分批接种于23只实验兔肝脏内，制成肝癌模型。将实验兔随机分为5组：对照组3只（A组），不行RFA治疗；余下20只行超声引导下RFA治疗，射频范围为肿瘤最大半径的2/3，人为造成肿瘤组织残余。按治疗后不同时间分为射频后0h组（B组）5只；射频后1w组（C组）5只；射频后2w组（D组）5只；射频后4w组（E组）5只。各组经超声检查后将实验兔处死，切取标本，采用免疫组织化学法观察A组肿瘤及B-E组残余肿瘤组织中IGF-II及PCNA的表达情况。结果：1．造模结果:接种后2-4w行超声检查，23只实验兔均种植成功。超声显示肝脏种植区圆形或类圆形低回声肿物，内回声均匀，大小1.5±0.5cm，边界清，无包膜回声，血流丰富。A组经病理证实，接种成功率100%。 2．RFA术后超声结果：术后0h肿瘤损毁区为不规则汽化强回声，边界不清，形态不规则，彩色多普勒（CDF）及能量多普勒（CDE）:无法测到血流信号。术后1w、2w、4w肿瘤大小与术前相似，治疗区呈不规则高回声，中心区可见少量不规则无回声；术后1w、2w CDF及CDE无明显血流信号，术后4w CDE部分残癌周围可见血流信号。 3．HE染色结果：A组瘤体及B-E组残癌细胞核大深染，异型性明显，呈巢状分布。射频区凝固坏死，可见核固缩、核碎裂、核溶解，周围见大量红染组织，淋巴细胞浸润，周围形成1-3mm炎症反应带；射频1w后开始出现液化，2w液化坏死范围扩大，炎症反应带纤维化。 4．免疫组织化学结果：A组IGF-II、PCNA呈高表达，分别为（86.3±12.5）、（65.37±13.36）；B组IGF-II、PCNA在残癌组织中表达阳性率为（78.9±13.3）、（50.78±13.98）；C组IGF-II、PCNA在残癌组织中表达阳性率为（47.2±10.2）、（35.63±10.20）；D组IGF-II、PCNA在残癌组织中表达阳性率为（18.9±4.3）、（17.80±5.17）；E组IGF-II、PCNA在残癌组织中表达阳性率为（10.6±1.89）、（9.91±1.63）。B-E组IGF-II、PCNA在消融区均低表达。结论:射频消融治疗可使兔肝VX2肿瘤残余组织中癌细胞失去活性，从而起到进一步的治疗作用。
[Abstract]:Objective: To observe the blood flow classification and blood flow of liver cancer, and to explore the relationship between the expression of hypoxia inducible factor -1 alpha (hypoxiainducible factor-1alpha, HIF-1 a) and microvascular density (microvessel density, MVD) in the liver cancer tissues and the relationship between the two.
Methods: 60 cases of primary liver cancer confirmed by pathology were examined by two-dimensional and color Doppler ultrasonography before operation, and the blood flow classification was carried out. The expression of HIF-1 alpha and MVD in liver cancer tissues was detected by immunohistochemistry.
Results: the ultrasound blood flow classification of liver cancer was I grade HIF-1a, MVD expression rate was (64.37 + 3.02), (39.32 + 3.71), blood flow classification was II grade HIF-1a, MVD expression rate was (79.94 + 2.97), (62.25 + 3.80), III grade HIF-1a, MVD expression rate was (92.61 + 3.10), (85.16 + 2.33), HIF-1 alpha, MVD phase of ultrasonic liver cancer When the blood flow classification was III grade, the expression rate of MVD was the highest, which was 92.61% + 3.10,85.16% + 2.33 respectively. The expression rate of HIF-1a and MVD in normal liver tissues was 12.28% + 7.21,11.33% + 7.21 respectively. The two groups had significant difference (P0.05).
Conclusion: the expression of HIF-1a is closely related to the formation of neovascularization of liver cancer. The higher expression of MVD, the higher the expression of HIF-1 a, the more rich the color Doppler shows, the ultrasound flow classification of liver cancer can provide a theoretical basis for the clinical diagnosis and treatment of liver cancer.
Objective: To study the expression of insulin growth factor -II (Insulin-like growth factor, IGF-II) and proliferating cell nuclear antigen (proliferating cell nuclear) in the residual cancer of rabbit liver VX2 tumor (Radiofrequency Ablation, RFA).
Methods: 28 rabbits and 3 rabbits with VX2 tumor bearing only were used as passage rabbits, and the remaining 23 rabbits were implanted with intrahepatic tumors under ultrasound guidance to establish a VX2 liver cancer model. The tumor tissue surrounding the tumor was harvested from the tumor rabbit. 2 rabbits were taken randomly from the tumor rabbits. After local anesthesia, the scalpel broke the skin, and the tumor block was sent to the subcutaneous and passages with tweezers. Under the guidance of ultrasonic guidance, the VX2 tumor particles were inoculated into 23 experimental rabbits' liver and made into a liver cancer model. The experimental rabbits were randomly divided into 5 groups: 3 control group (group A) and no RFA treatment; the remaining 20 were treated by ultrasound guided RFA, the range of radiofrequency was 2/3 of the maximum radius of the tumor. 5 rats were divided into group 0h (group B) after radiofrequency, 5 in group 1W (group C) after radiofrequency, 5 in group 2W (group D) after radiofrequency, 5 in group 4W (group E) after radiofrequency. After ultrasound examination, the rabbits were killed and specimens were cut and specimens were cut, and the expression of IGF-II and PCNA in group A tumor and B-E group was observed by immunohistochemistry.
Results: 1. the results were as follows: 2-4w ultrasound examination after inoculation, 23 rabbits were planted successfully. Ultrasound showed round or circular hypoechoic tumor in the liver planting area. The internal echo was uniform, the size was 1.5 + 0.5cm, the boundary was clear, no envelope echo, the blood flow rich.A group was confirmed by pathology, and the success rate of grafting was 100%.
2.RFA postoperative ultrasound results: postoperative 0h tumor damaged area was irregular vaporized strong echo, irregular boundary, irregular shape, color Doppler (CDF) and energy Doppler (CDE): no blood flow signal was detected. Postoperative 1W, 2W, 4W tumor size was similar to preoperative, irregular hyperechoic in the treatment area, and a small amount of irregular anechoic in the center area; 1 after operation. W, 2W CDF and CDE had no obvious blood flow signals. Blood flow signals were found around 4W CDE partial residual tumor after operation.
3.HE staining results: the cell nuclei of tumor body and group B-E in group A were deeply dyed, with obvious heteromorphosis and nesting distribution. The radiofrequency region solidified and necrotic, visible nuclear condensation, nuclear fragmentation, nuclear disintegration, a large number of red dyed tissues, lymphocytic infiltration, and peripheral formation of 1-3mm inflammatory zone; 1W began to liquefy after 1W, and 2W liquefaction necrosis expanded, inflammation expanded. The disease reacts with fibrosis.
4. immunohistochemical results: A group IGF-II, PCNA showed high expression, respectively (86.3 + 12.5), (65.37 + 13.36), IGF-II, PCNA in the B group was (78.9 + 13.3), (50.78 + 13.98), C group IGF-II, PCNA in the residual cancer tissues (47.2 + 10.2), (35.63 + 10.20), D IGF-II, PCNA in the residual cancer tissue expression. The positive rate was (18.9 + 4.3), (17.80 + 5.17), and the positive rate of IGF-II in E group was (10.6 + 1.89), (9.91 + 1.63).B-E group IGF-II, and PCNA was low expression in the ablation area.
Conclusion: radiofrequency ablation can make the cancer cells lose their activity in the remnant tissues of VX2 tumor of rabbit liver, thereby playing a further therapeutic role.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R445.1;R735.7

【参考文献】