| Breast cancer is a common female malignancy in a highlyheterogeneous, our current breast cancer incidence is2%to3%annualgrowth rate. Breast cancer MDA-MB-231cell lineis triple-negative breastcancer, and is a subtype of Basal B cells from pleuraleffusion.MDA-MB-231is a metastatic adenocarcinoma. Due to its highinvasive and high transfer characteristics, MDA-MB-231is currently theworst prognosis subtype of breast cancer.Nude mice appeared in1966, the two transcription factors FoxN1and fork-head box N1encoding a forkheadfamily proteins, and these twogene mutations generated nude mice. These two genes are mainlyexpressed in the thymic epithelium, wherehas obviously keratosis andhair follicles. FoxN1sequence and functions are highly conserved inrodents and humans. In mice, rats and humans, FoxN1natural mutationscan lead to thymic hypoplasiaand glabrous skin. In humans, FoxN1mutations in humans can cause hair loss and nail dystrophy, and lead toserious infections associated with primary T-cells deficiency. But in mice,the FoxN1mutant results in generating nude mice, due to its T celldevelopment immune deficiency and hair follicle development failure hasbeen widely used as an experimental model in tumor, immunology andtransplantation.Small animal ultrasound and in vivo fluorescence imaging are two experimental methods which are widely used in animalexperiments.Different to pathological process in which analysis requiresexperimental animals to be sacrificed for specific test,the first twoexperimental techniques can be the case in the absence of trauma on thedevelopment of animal models for testing, and because the same mice hasbeen used before and after the experiment, individual differences can bereduced, thereby reducing experimental error. However, there aredifferences between the two experimental means, such as small animalultrasound can use different frequency probes to detect animals indifferent organs, but fluorescence imaging cannot locate, but in vivofluorescence imaging can visually observe fluorescence expression sites,without a step by step investigation.In this study, breast cancer cells MDA-MB-231that expressed ofGFP were transplanted into nude mice, and successfully establishedbreast cancer transplanted tumor model. Then using small animalultrasound imaging and in vivo fluorescence imaging to monitortransplanted tumor occurrence and development process, andcomparingthe different role ofbetween small animal ultrasound imagingand in vivo fluorescence imaging in this process. Meanwhile,we appliedpathological analysis in transplanted tumors to compare the differenteffects between small animal ultrasound imaging and HE staining inbreast tumor transplantation model.Objective:1. Established breast cancertransplanted tumor model in nude mice.2. Observe the effect of small animal ultrasound imaging in breastcancer transplanted tumor study.3. Observe the role of in vivo fluorescence imaging in breast cancertransplanted tumor study. Method:1. Stably expressing GFP breast cancer cells MDA-MB-231wereinoculated into nude mice at its right breast, and establish breast cancertransplanted tumor in nude mice.2. Nude mice were anesthetized with0.1g/ml sodium pentobarbital,use a small animal ultrasound imager to monitor the development andprogression of breast cancer transplanted tumor in nude mice.3. Nude mice were anesthetized with0.1g/ml sodium pentobarbital,use a small animal in vivo fluorescence imaging to monitor thedevelopment and progression of breast cancer transplanted tumor in nudemice.4. Tumors in nude mice were sacrificed on day28, take nude mousetransplanted tumor for HE staining.Result:1.Successfully constructed breast cancer transplanted tumor modelin nude mice.2. HE staining:Transplanted tumor wasmesenchymal tumor, mostlyconsisted of fibrous cells, relatively few cancer cells, cancer cells areirregular shape, split-phase, nucleiswere large and deeply stained, cancercells has a big nuclear-cytoplasmic ratio, showing a low degree ofdifferentiation features; small blood vessel formation were seen.3. Small animal ultrasound imaging to monitor the development andprogression of breast cancer transplanted tumor.(1) in day3of tumor inoculation,no tumor were detected;(2)in day7of tumor inoculation, on the right side of the nude mice’schest, ultrasound shows uneven hypoechoic area of2.37mm2, longdiameter of2.44mm, short diameter of0.28mm, it was ill-defined;(3) in day14of tumor inoculation, on the right side of the nudemice’s chest, ultrasound shows uneven hypoechoic area of6.62mm2, longdiameter of3.68mm, short diameter of1.66mm, itwas ill-defined, and uneven thickness of the halowere seen at the surrounding, punctatehyperechoics were seen in it;(4) in day21of tumor inoculation, on the right side of the nudemice’s chest, ultrasound shows uneven hypoechoic area of21.99mm2,long diameter of7.59mm, short diameter of3.33mm, itwas ill-defined,hyperechoics were seen in it;(5) in day28of tumor inoculation, on the right side of the nudemice’s chest, ultrasound shows uneven hypoechoic area of52.00mm2,long diameter of9.28mm, short diameter of8.06mm, itwas ill-defined,and uneven thickness of the halowere seen at thesurrounding,morehyperechoics were seen in it;4.in tumor inoculation day, namely day0, GFP expression can beobserved by in vivo fluorescence imaging, but the expression offluorescentwere not checked on day7, at day14, high intensityfluorescent expression were observed, and gradually increased over time.Conclusion:1. Successfully constructed breast cancer transplanted tumor modelin nude mice.2. Fluorescence Imaging has highly sensitivity in monitoring tumor early development and progression.3. Small animal ultrasound has highly sensitivity and accuracy in monitoring the middle and advanced stage of tumor development and progression, and more convenient in operation.4. Relative to pathology analysis, small animal ultrasound has advantage of real-time monitoring to ensure the continuity and consistency of live animal experiments, and reduce experimentalerror, shows the unique advantages in animal experiments. |
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