| Objective:The subcutaneous and orthotopic transplantation model of LNCaP prostate cancer cells in nude mice was established.The influence of clinical features on the detection rate of human prostate cancer was analyzed through transrectal ultrasound-guided transperineal prostate puncture biopsy.Methods:1.LNCaP cells were revived,the state of cell growth throughout time were observed,and the methods of cell resuspension was used to deal with LNCaP cells.2.LNCaP cells were implanted subcutaneously into BALB/c nude mice to construct the prostate cancer model.Ultrasonic shear wave elastography(SWE)was used to measure the velocity of wave propagation in the tumor and analyze the pathological changes in the tumor tissue;3.LNCaP cells were injected into the prostate of BALB/c nude mice to construct an orthotopic transplantation model;The tumor generation and volume changes were monitored by ultrasound imaging.The blood perfusion in the tumor was examined by contrast-enhanced ultrasound and the region of interest(ROI)generation time intensity curve(TIC)was delineated.The arrival time(Atm),time to peak(Tto Pk)and area under the curve(Au C)of contrast agent entering tumor were automatically generated and recorded by the instrument.The expression of androgen receptor(AR)was analyzed by pathological examination,and the expression of platelet endothelial cell adhesion molecule(CD31)and vascular endothelial growth factor A(VEGF-A)in tumor tissues was detected by immunohistochemical staining.Immunohistochemical results were analyzed by Image J software and the average optical density(AOD)of CD31 and VEGF-A antibody staining was calculated;Finally,Pearson correlation analysis was performed on the contrast-enhanced ultrasound parameters Atm,Tto Pk,Au C and tumor vascular immunofluorescence index AOD using Prism 9.0 statistical software.4.A comparative retrospectively study were conducted.A total of 84 patients were referred to biopsy in this research.According to the gold standard pathological results,the patients were divided into positive(n=39)and negative groups(n=45),and clinicopathological factors were compared between the groups.Independent sample T-test was used to analyze the groups difference of age and the number of puncture needles,and Mann-Whitney test was used to analyze other clinical data.Receiver operating characteristic(ROC)curve was used to analyze the diagnostic efficacy of clinical characteristics on transperineal prostate biopsy guided by transrectal ultrasound Results:1.LNCaP cells were revived in 24 hours later,many cells gathered into clusters,closely connected with each other,and rod-shaped branches grew outward at the edge of cells;After 48 hours,the cells were clustered,and there were fewer rod-shaped cell branches;After 72 hours,the number of cells was generally small,and the fusion of lamellae cell clusters could be seen;After revived human prostate cancer LNCaP cells for 12 hours,single cell suspension was observed,and a few cells were in adherent state;After 48 hours,more rod-shaped LNCaP cells adhered to the wall;After 72 hours,the cells were connected with each other by synapses,and LNCaP cells were completely bifurcated,extending outward by synapses;2.On the 85th day of subcutaneous tumor take rate was 83.3%(5/6)in nude mice.On the 33rd day,the tumor volume was 313.28±181.77mm~3,and on the 75th day,the tumor volume was 518.50±186.88mm~3;In the methods of CDFI,there were spot and strip blood flow signals behind the tumor edge,but no obvious blood flow signals inside the tumor;Under the ultrasonic elastic imaging,the propagation velocity of shear wave in the subcutaneous transplanted tumor was 2.40±0.33m/s;3.On the 50th day,the orthotopic tumor take rate was 71.4%(15/21)and the average volume was 288.81±76.49mm~3;Contrast-enhanced ultrasound curve analysis showed that Atm was 1.59±0.65s,Tto Pk was 11.59±3.68s,Au C was 1933.91±454.38d B;Surgical anatomy found that the tumor was oval in shape,immunohistochemical staining of tumor tissue showed that the androgen receptor(AR)staining in tumor cells was strongly positive,AR was located in the nucleus;the positive expression of CD31and VEGF-A by immunohistochemical staining,and Image J software analysis showed that AOD of CD31=0.39±0.06 and VEGF-A=0.45±0.05,Prism statistical analysis showed that the correlation between VEGF-A and Atm,Ttopk,Au C was r=-0.9308(P<0.0001),r=-0.8782(P<0.0001),r=0.8675(P<0.0001);the correlation between CD31 and Atm,Ttopk,Au C was r=-0.6783(P=0.0002),r=-0.3601(P=0.0180),r=0.3097(P=0.0312).4.The age of 84 patients with prostate biopsy was(67.64±8.57)years.After ultrasound measurement,the median of the total volume of prostate was 47.08mm~3,and the median of the volume of prostate was 23.45mm~3;There was no statistically significant difference in the detection rate of prostate cancer between the total volume of prostate and prostate-specific antigen(PSA)concentrations(P>0.05).There was statistically significant difference in the detection rate of prostate cancer between the age of patients with prostate cancer,the area of prostate transitional zone,the number of puncture needles,ultrasound elasticity score and prostate-specific antigen density(PSAD)(P<0.05);The analysis of clinical characteristics by transrectal ultrasound-guided transperineal prostate puncture showed that the sensitivity and specificity of age for the diagnosis of prostate cancer were 61.54%and 75.56%.ROC curve showed that the area under the curve was 0.705(95%CI:0.593～0.818,Z=3.591,P=0.0003),the maximum value of Yoden index is 0.3709;The sensitivity and specificity of needle number for the diagnosis of prostate cancer were 35.90%and 86.67%.ROC curve showed that the area under the curve was 0.613(95%CI:0.493～0.733,Z=1.854,P=0.064),and the maximum Yodon index was 0.2256.The sensitivity and specificity of total prostate volume for the diagnosis of prostate cancer were 64.10%and 64.44%.ROC curve showed that the area under the curve was 0.607(95%CI:0.482～0.733,Z=1.680,P=0.093),and the maximum Yoden index was 0.2855.The sensitivity and specificity of PSAD for the diagnosis of clinical prostate cancer were 92.31%and28.89%.ROC curve showed that the area under the curve was 0.606(95%CI:0.791～0.984,Z=2.622,P=0.009),and the maximum Yodon index was 0.2120.The sensitivity and specificity of the volume of the prostate transitional zone for the diagnosis of prostate cancer were 58.06%and 75.56%.ROC curve showed that the area under the curve was 0.640(95%CI:0.507～0.773,Z=2.064,P=0.039),and the maximum Yodon index was 0.3362.The sensitivity and specificity of ultrasonic elasticity score for the diagnosis of prostate cancer were 57.89%and 85.71%.ROC curve showed that the area under the curve was 0.734(95%CI:0.605～0.863,Z=3.549,P=0.0004),and the maximum value of Yodon index was 0.4361.Conclusion:1.LNCaP cell growth after cultured with 12h as adhere cell.At 72h,the cells branched and extended outward with multiple antennae,which could be improved by re-suspension of cell suspension when contact inhibition of cell aggregation occurred;2.The tumor take rate in the subcutaneous transplantation model of BALB/c nude mice by the methods of LNCaP cell suspension+matrigel is higher;3.Ultrasound is useful monitor tool that the pathological changes of the internal tissue of the subcutaneous transplanted tumor in Balb/c nude mice;4.The injection methods of LNCaP cell suspension+matrigel is successfully construct the orthotopic transplantation model of prostate cancer in nude mice of BALB/c,and improve the tumor take rate of the model;5.Ultrasound is useful monitoring tool of tumor growth in BALB/c nude mice prostate cancer orthotopic xenograft models,and contrast-enhanced ultrasound for accurate preclinical evaluation of androgen dependent prostate cancer involving angiogenesis and antiangiogenesis therapies;6.The monitoring of clinical characteristics with age,prostate volume,PSAD,prostate transition zone volume and ultrasonic elasticity score might improve the diagnostic capacity of transrectal ultrasound-guided transperineal prostate biopsy. |
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