不同方法建立小鼠肝癌原位移植瘤模型差异性的探讨

Different transplantation models of hepatocellular carcinoma in mice

目的 比较肿瘤细胞注射和肿瘤组织块移植方法建立小鼠肝癌原位模型的差异,为小鼠肝癌原位模型的建立提供技术参考。方法 取健康雄性KM小鼠,随机分为4组:A组注射小鼠肝癌H22细胞;B组注射腹水型肝癌H22细胞;C组采用小鼠肝癌组织块移植法;D组肝部注射生理盐水作为假手术组。定期观察各组小鼠活动情况、体重变化。记录4组小鼠的生存时间。观察各组小鼠肝部成瘤状况、肿瘤程度、与腹腔脏器粘连程度和转移情况,并进行B超成像,检测血清中甲胎蛋白(alpha-fetoprotein, AFP)、异常凝血酶原(des-gamma-carboxyprothrombin, DCP)的浓度及苏木素-伊红(HE)染色观察肝组织病理学变化。结果 A组、B组、C组小鼠造模操作时间分别为(3.36±0.44)min、(3.30±0.41)min、(5.68±0.65)min。建模第25天,A组、B组、C组小鼠成瘤率均100.0%,A组腹腔重度粘连率为40.0%,B组腹腔重度粘连率为60.0%,C组与D组均未出现重度腹腔粘连。A组、 B组、C组小鼠腹水出现率分别为40.0%、100.0%、0.0%;腹壁瘤出现率分别为30.0%、60.0%、0.0%。B组小鼠还存在肝转移情况(40.0%)。B超成像、血清AFP和DCP水平检测、组织病理学检查结果显示,与D组小鼠相比,A组、B组、C组小鼠肝边缘均不光滑,回声欠均匀,可见稍低回声包块;保持AFP、DCP高分泌;存在大量炎症细胞与肿瘤细胞。结论 在第25天时,3种方法均可以建立肝癌原位移植模型,其中,相较于肝癌组织块移植,肿瘤细胞注射具有操作简单、肝内存在多个转移结节的特点。而肝癌组织块移植相较于肿瘤细胞注射,则具有对腹腔等脏器影响较小等特点。

Objective To explore the otherness of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish orthotopic implantation models of hepatocellular carcinoma in mice,and to provide a technical reference for the establishment of an orthotopic implantation model.Methods Healthy KM mice were divided into four groups:group A,direct injection of H22 cells;group B,direct injection of H22 ascitic cells;group C,transplantation of tissues;and group D,direct injection of saline.Activity and weight changes were observed regularly in each group and survival times were recorded.Liver tumor formation,tumor size,abdominal organ adhesion degree,and metastasis were observed in all groups.B-ultrasound imaging was performed,concentrations of alpha fetoprotein(AFP)and abnormal prothrombin(DCP)were detected,and liver histopathological changes were detected by hematoxylin and eosin staining.Results Mice molding operation time in groups A,B,and C were(3.36±0.44)min,(3.30±0.41)min,and(5.68±0.65)min,respectively.After modeling for 25 days,the rates of model formation in groups A,B,and C were all 100.0%.Severe abdominal adhesions occurred in 40.0%of mice in group A and 60.0%in group B,but in no mice in group C or D.Ascites occurred in 40.0%,100.0%,and 0.0%and abdominal wall tumors in 30.0%,60.0%,and 0.0%of mice in groups A,B,and C,respectively,while 40.0%of mice in group B also had liver metastasis.B-ultrasound imaging,detection of serum AFP and DCP levels,and histopathological result showed smooth liver margins,uneven echo and slightly lower echo mass,maintained high AFP and DCP secretion,and large numbers of inflammatory cells and tumor cells in mice in groups A,B,and C.Conclusions At day 25,all three methods can thus be used to establish orthotopic transplantation models of HCC.Among these,inj ection of cell suspensions demonstrated the advantage of simplicity in operation and the presence of multiple metastatic nodules within the liver,compared to transplantation of tumor tissue.Conversely,transplantation of tumor tissue showed the advantage of causing less impact on the abdomen and other organs when compared to inj ection of cell suspensions.