Résumé
Objective@#To investigate expression of nucleolar protein 14(NOP14) and CD31 in pancreatic cancer mouse model and its correlation with tumor progression.@*Methods@#Clinicopathological data of 5 patients with pathologically confirmed pancreatic ductal adenocarcinoma(PDAC) and hepatic metastasis between January 2013 and December 2015 was collected in Department of General Surgery, Peking Union Medical College Hospital. Immunohistochemistry staining was employed to detect the expression of NOP14 in matched primary PDAC and relevant metastasis.Pancreatic cancer cells with NOP14 stably knocked down were established by transfecting lentivirus with NOP14 targeted silencing RNA.The inhibition efficacy was detected by quantitative real time PCR and western blot.Microvascular density(MVD) in pancreatic cancer transplantation mouse model was determined by CD31 immunohistochemistry staining analysis and correlated with NOP14 expression and tumor progression.@*Results@#NOP14 had a significant higher expression in liver metastasis than primary pancreatic adenocarcinoma (2.09±0.45 vs. 1.31±0.27, P=0.028). NOP14 was knocked down 86 percent on mRNA level determined by qPCR and 78 percents on protein level detected by western blot. MVD was significantly decreased in NOP14-inhibited tumor from both pancreatic cancer cells subcutaneously and orthotopically grafted tumor mouse model with the value of 61.40±13.85 vs. 85.53±14.59 (P=0.041) and 38.33±10.91 vs. 59.33±15.37(P =0.037), respectively. Besides, MVD was positively associated with tumor volume(r=0.842, P<0.01) and metastasis (r=0.726, P=0.008).@*Conclusion@#NOP14 presents higher expression in hepatic metastasis of pancreatic adenocarcinoma and might promote tumor progression by increasing microvascular density.
Résumé
Objective To investigate the impact of 5-fluorouracil (5-FU) and cisplatin on miR-449b expression in human hepatocellular carcinoma (HCC) and elucidate the molecular mechanism of 5-FU and cisplatin inhibiting the migration of HCC cells.Methods Real-time qPCR analysis was conducted to determine the expression of miR-449b in 50 HCC tissues.RT-PCR assay was performed to detect the expression of miR-449b in HCC cells with 5-FU and cisplatin treatment.The migration of HCC cells with the overexpression of miR-449b was determined by wound-healing assay;Rescue assay was employed to investigate the correlation between 5-FU & cisplatin, miR-449b and the migration capacity of HCC cells;The putative targets of miR-449b were predicted and validated using target prediction programs and immunoblots.Results The expression of miR-449b decreased in HCC tissues (P<0.0001).miR-449b expression increased in HCC cells upon the treatment of 5-FU and cisplatin (P<0.001).The overexpression of miR-449b inhibited the migration of HCC cells (P<0.001).Rescue assay revealed that inhibition of miR-449b to prevent 5-FU and cisplatin induction resulted in suppressed migration in SMMC7721 cells(P<0.05).Catenin-δ was a functional target of miR-449b.Conclusions 5-FU and cisplatin inhibit the migration of HCC cells at least partly via inducing the expression of miR-449b.
Résumé
Objective To investigate the expression profile of miRNAs up-regualted in human extrahepatic and intrahepatic cholangiocarcinoma tissues and probe the effect on cell growth of four of these miRNAs in QBC939 cell line.Methods Up-regulated miRNAs in extrahepatic or intrahepatic cholangiocarcinoma tissues were analyzed by using miRNA-microarray,which was confirmed by using miRNA Real-Time PCR analysis.Based on these findings,four of these up-regulated miNRAs were chosen to perform function investigation.The specific miRNA inhibitors were transfected into QBC939 cells,respectively,and cell proliferation assay was performed by using MTT.Results 12 miRNAs were up-regulated both in two types of cholangiocarcinoma tissues,28 miRNAs and 21 miRNAs were up-regulated in extrahepatic cholangiocarcinoma and intrahepatic cholangiocarcinoma,respectively.MiR-125b and miR-19a expression levels were increased about 3.7 and 3.6 fold,compared with the matched normal bile duct tissues (P＜0.05).MiR-92a and miR-205 expression was upregulated about 4.S- and 3.5-fold,compared with the matched normal bile duct tissues (P＜0.05).MiR-125b,miR19a,miR-21,and miR 378* were inhibited in QBC939 cells,which indicated a significant inhibitory effect on cell growth.The ratio of inhibition was 71％,72％,69％,and 76％(P＜0.05)at 36 h,61％,63％,60％,and 59％(P＜0.01) at 48 h,and 61％、56％、60％ and 59％(P＜0.05) at 60 h.Conclusion The miRNAs expression patterns in human extrahepatic and intrahepatic cholangiocarcinoma tissues are different and uo-regulated miRNAs act as oncomirs on cholangiocarcinoma cell growth.
Résumé
ObjectiveTo investigate the inhibitory effect of all-tram-retinoicacid (ATRA) on HCC cell growth and probe the potential molecular mechanism.MethodsHCC cell lines,HepG2 and SMMC-7721 were treated by ATRA and cell growth was analyzed by using MTT assay.The expression levels of miR-18a were evaluated in HepG2 and SMMC-7721,compared with the normal livers pool by using RealTime PCR analysis.Cell growth analysis by using MTT assay was performed on HepG2 and SMMC-7721 after transfection with anti-miR-18a.Rescued assay was designed to probe the mechanism of ATRA on cell growth by using ATRA with or without miR-18a mimic.ResultsHepG2 cell growth was suppressed about 74％ (P＜0.05,36 h),72％ (P＜0.01,48 h),and 67％ (P＜0.05,72 h) and SMMC-7721 cell growth was inhibited about 68％ (P＜0.05,48 h),and 64％ (P＜0.05,72 h) after treatment with ATRA,compared with the cells treated with Ethanol.MiR-18a expression was up-regulated in HepG2 and SMMC-7721 cell lines about 4.7- and 3.8-fold (P＜0.05),respectively.Endogenous miR-18a levels were down-regulated by ATRA about 67％ and 56％ (P＜0.05).The inhibitory effect of ATRA on HCC cell growth was reversed about 1.2-fold (P＜0.05,48 h) by overexpression of miR-18a in HepG2 cells and cell growth of SMMC-7721 was enhanced about 1.25- and 1.2-fold (P＜0.05,24 and 48 h) with ectopic expression of miR-18a.ConclusionHCC cells growth is suppressed by ATRA through miR-18a mediated network.
Résumé
【Objective】 To study the location effect of the Mc Ab G9 to human esophageal carcinoma in tumor bearing nude mice. 【Methods】 125 I-G9 was prepared by Chloramine-T method. The distribution of 125 I-G9 was detected at different time (24, 48, and 72 h) after peritoneal injecti on. The percentage of the injected dose per gram of tissue(%ID/g) and the ratio of Tumor/non-Tumor were calculated. 【Results】 The distribution of 125 I -G9 in tumor at the third day was showed by autoradiography obviously higher th an in other organ/tissue (except blood) and the highest is at the 48 h. The T/NT values are 2-7. The autoradiography indicated that radioactivity concentrated in tumor tissue. The concentration in tumor edge is more obvious than in the ce nter. 【Conclusion】 125 I-G9 has a considerable targeting activity and can well locate in esophageal carcinoma tissue in tumor bearing nude mice.
Résumé
In this study, immunotoxin (IT) was prepared by conjugating BAC5 and CT with SPDP. The effects of IT on NPC and its mechanisms were explored using double labeled with radioactive nuclides, immunography and electron microscope technique in vivo and in vitro. The specific concentration of BAC5 in the tumor area showed. The radioactivity rate of tumor/nontumor (T/NT) was up to 10.26. IT had cytotoxic effects both on the cultured CNE-2 cell line and tumor multicell spheroides. In vivo, the preliminary result indicated that IT also had a inhibitory action on the nude mice models bearing human NPC (Reported in another article). Under electron microscope, the necrosis and apoptosis of tumor cells were found. The membranes of most tumor cells were found intacted not or corrosined, some of them had the character of apoptosis, including reduce of tumor cells membrane villi, condensation of cytoplasm and pyknosis or cleavage of nuclear. There were many of apoptosis bodies, which were occasionally phagocytosed by tumor cells. The infiltration of immunocytoes in tumor tissue could be seen. The results indicated that BAC5 can specifically combine with NPC cells and BAC5-CT has the inhibitory effect on NPC in vitro and in vivo, mechanism of which may be related to the effects that ‘warhead' CT dissolute the membrane of tumor cells directly, or/and IT promote the infiltration of immunocytoes so as to induce the apoptosis of tumor cell.
Résumé
Objective To evaluate the feasibility of ~ 125 Iodine -labeled IL-2 scintigraphic methods for the imaging of lymphocyte infiltration of the Langerhans′ islets. Methods ~ 125 I-IL2 was injected intravenously to pre-diabetic female NOD mice (aged 16-17 weeks)and female Balb/c mice as control. Animals were sacrificed at different time points and the radioactivity in different organs was measured.~ 131 I-IL-2 was injected iv to 3 pre-diabetic female NOD mice, and pancreas was imaged through SPECT after 30 minutes injection. Results The mean radio activities in the pancreas of NOD mice were significantly higher as compared to those of Balb/c mice at time points from 30 to 120 minutes (P
Résumé
AIM: To investigate the effect of 188Re labeled monoclonal antibody on prostatic specific membrane antigen 7E11C5.3,radioimmunotherapy for the treatment of human prostate cancer cell line LNCaP in vitro.METHODS: 188Re-7E11C5.3 was prepared by direct 2-mercaptoethanol reduction method.Labeling efficiency and radiochemical purity was measured by paper chromatography.Immunoreactive fraction was determined by linear extrapolation.Cytotoxicity to LNCaP cells was determined by MTT assay.RESULTS: The labeling yield of 188Re-7E11C5.3 was(93.16?2.18)%,the radiochemical purity was(95.62?0.48)%,and the immunoreactive fraction was(74.86?1.86)%.The inhibitory effect of 188Re-7E11C5.3 on cell proliferation of LNCaP cells was significantly higher than that of 188Re-mIgG or 188ReO-4.The 50% inhibitory doses(IC50) of 188Re-7E11C5.3,188Re-mIgG,and 188ReO-4 were(23.38?3.73)?107 Bq/L,(59.21?8.02)?107 Bq/L and(68.89?10.91)?107 Bq/L,respectively.CONCLUSION: 188Re-7E11C5.3 can effectively inhibit the growth of in vitro cultured prostate cancer cells and shows much potential for prostate cancer radioimmunotherapy.