The Study Of Contrast-enhanced Ultrasound And Its Biological Character In Cervical Transplanted Carcinoma

ObjectiveCervical carcinoma is a common malignant tumor.To establish an animal model of cervical cancer in immunocompetent rat and to study its biological character.Based on pervious study,the purpose of this study is to found Hela mice tumor xenograft model and study the,through which to provide a way to prevent and cure the cervical cancer in clinic.To explore the regularity of the ultrasonographic dynamic changes such as the size,echo-intensity and edge echo-pattern of cervical tumor during its growth. To offer the animal experimental proves of the preclinical phase for the early diagnosis and treatment of cervical carcinoma.To investigate P38MAPK protein expression and its clinical significance in the cervical transplanted carcinoma of rat.In the meanwhile, to assess the blood perfusion of tumors in rabbit muscles with perfluorocarbon contrast flow imaging and high-frequency ultrasound.MethodsPart one:Hela was frozen in liquid nitrogen were revived in routine method and inoculated in RPMI-1640 Blended with 10%foetus cattle serum.HELA cells 2×10~7,2×10~6,2×10~5 was injected intraperitoneally,under skin,and vein.Human cervical cancer Hela cells were implanted into the mice to get the tumor model of HELA cell line.The transplanted tumors were observed every two days.The ultrasonographic observation was performed on the 7,10,14,21 and 30 day after implantation.The survivals were put to death and necropsy.The section preparations were stained with HE method.Part two:Hela was frozen in liquid nitrogen were revived in routine method and inoculated in RPMI-1640 Blended with 10%foetus cattle serum.2×10~5 was injected under skin. Immunohistochemical assay and Westem Blot was performed to study the expression and location of P38.Subcutaneously injected with PBS as a control group in 5 mice.Part Three:Ten rabbits model with cervical transplanted tumors in muscle were examined, and they were assessed by high-frequency color Doppler ultrasound every week for ten weeks.SonoUve was administered respectively to ten rabbits through ear veins at 0.2, 0.4,0.6 ml/kg,and then imaged with second harmonic imaging.After intravenous administration of contrast agent,Color Doppler imaging or Power Doppler imaging were used to evaluate the blood perfusion of tumors in muscle.ResultsHela cells grew progressively after injected into rats intraperitioneally,formed gross peritoneal dissemination and malignant hemorrhagicascit-es in a manner typical for human cervical carcinomas.Animal survival time decreased significantly with large Hela burden(p＜0.01).Pathological analysis and tissue culture confirmed the existence of the carcinoma tissue.The implant,The eho-intensity of the tumor can be divided into three patterns such as hyperecho,isoecho and hypoecho.The observed rate of abnormal vessels in the tumor increased as the implantation time elongated and the tumor enlarged.The observed time of arterial blood was earlier than that of venous blood.The effective factors of histopathology mostly were necrosis,fiber and sinusoid. In two -dimensional images,tumors were small and poor echo one week after implantation.Necrosis and liquefaction appeared could be found later at the 6-7 week cavum formed.Color Doppler flowing showed dot,branch,or line flow signals and the degree of blood abundance was from 0-Ⅲgrades,while PI and RI between phrases had no significant difference,there were significant improvements in detecting neovascularity of tumor in muscle at all contrast agent dosage levels and the intensity lasted a long period.P38 was mainly expressed in the cell cytoplasm and none nuclei of cervical cells and neutrophil and lymphcytes.Positive rate of p38MAPK protein expression was 70%in carcinoma tissues,33.1%in its paraneoplastic tissue,and there is a significant correlation between the expression of P38 with the stages of cervical cancer,histology grade,and local lymph node ivolvement,but not with time,site or size of tumor.ConclusionsA positive cervical cancer model has been successfully established in rats which have immunocompetent function that simulates preferably the biological behavior of human spontaneous cervical carcinoma.And is an optimal animal model for cervical cancer immunological therapy research.The ultrasonographic signs such as the size, echo -intensity,edge echo-pattern and blood flow changes of tumor during its growth were coincident to clinical cervical tumor.The gradual growth and gradual behavior of ultrasonography were essentially coincident to the defintive characteristics of tumor invasion.This animal model can be used to observe the malignant biological behaviors. The sizes of most tumors were less than 3cm,therefore the dynamic observation of the ultrasonographic roution parameters is useful to the diagnosis and odentification of the clinical little tumor.P38 is overexpressed in cervical cancer cells.The expression of P38 has a significant correlation with the stages of cervical cancer,p38 may play an important role in the occurrence,development and metastasis of human cervical carcinoma.The contrast agent is able to detect clearly the tumor vessels which cannot be shown with conventional color flow imaging.Moreover,the perfluorocarbon contrast agent can maintain the visuality of the vessels for a longer period.