OBJECTIVETo construct two recombinant plasmids of mdr1 and mcl1 shRNA, and to investigate their reversal effect on drug resistance in K562 adriamycin resistant cell lines (K562/A02).

METHODSTwo oligonucleotides of mdr1 and mcl1 gene were designed referring to that of GenBank, double-stranded DNA was derived through annealing, and cloned into pRNAT vector digested by two restricted endoenzymes. K562/A02 cells were transfected with the recombinant plasmids. The mdr1 mRNA expression and its protein product P-glycoprotein (P-gp) were detected by RT-PCR and flow cytometry. The expression of mcl1 gene was detected by RT-PCR. 50% inhibition concentration (IC50) of adriamycin (ADM) on K562/A02 cells was determined by MTT method. Cells apoptosis was analyzed by flow cytometry.

RESULTSComparing with K562/A02 cells, the shRNA of mdrl or mcl1 gene in vitro can remarkably increase the sensitivity of K562/A02 to adriamycin, down-regulate mdr1 or mcl1 gene expression, increase the K562/A02 cells apoptosis rates induced by adriamycin. Cotransfection of mdrl and mcl1 genes shRNA can also down-regulate the expression of their gene, more remarkably increase the sensitivity and apoptosis of K562/ A02 to adriamycin.

CONCLUSIONTransfection of mdrl or mcl1 gene shRNA can promote the sensitivity of K562/A02 to adriamycin and cotransfection of the two shRNA can more remarkably do so. The mel1 gene might be involved in adriamycin resistant in K562/A02 cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Apoptosis ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Flow Cytometry ; Gene Expression ; Humans ; K562 Cells ; Myeloid Cell Leukemia Sequence 1 Protein ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo explore the role of Notch signaling in human breast cancers, the expression of Notch1 and its ligand JAG1 in human breast cancers and their relationships with clinical stages of breast cancers were analyzed.

METHODSRT-PCR was used to detect the expression of Notch1 and JAG1 in 62 breast cancer specimens and 22 normal breast tissues at the margin of tumor sections, and the statistical difference of expression rates and standardized coefficient between the two groups were analyzed. To compare the expression intensity of Notch1 and JAG1 at different development stages of the illness and at different stages with or without axillary node metastasis.

RESULTSThe expression rate and standardized coefficient of Notch1 in human breast cancers were significantly higher than those of normal breast tissues at the margin of tumor sections. The expression rate of JAG1 in human breast cancers was 15%, while JAG1 was not detected in normal breast tissues at the margin of tumor sections. The standardized coefficient of Notch1 in cases with axillary node metastasis was significantly higher than that in cases without axillary node metastasis. The standardized coefficient of Notch1 at stage I was significantly lower than that at stage II, and stage II was significantly higher than stage III. There was no statistically significant difference between stage I and stage III.

CONCLUSIONNotch1 and JAG1 are highly expressed in human breast cancers, indicating that the aberrant expression and activation of Notch1 may be related with tumorigenesis of human breast cancer. Notch1 may play different roles at different developmentl stages of human breast cancer.

Adult ; Aged ; Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; pathology ; Calcium-Binding Proteins ; genetics ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jagged-1 Protein ; Lymphatic Metastasis ; Membrane Proteins ; genetics ; Middle Aged ; Neoplasm Staging ; Receptor, Notch1 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Serrate-Jagged Proteins ; Signal Transduction ; genetics

Objective To investigate the safety and efficacy of 177Lu-prostate specific membrane antigen (PSMA)-617 in the treatment of metastatic castration-resistant prostate cancer (mCRPC).Methods From August 2017 to September 2018,11 patients(average age 70.6 years) with mCRPC who underwent 177Lu-PSMA-617 therapy in Nanjing First Hospital were studied.All patients underwent 68Ga-PSMA-11 PET/CT before therapy to assess the tumor radioactive uptake.Blood routine examination and renal function test results were documented before and after therapy to assess the safety.The efficacy was reflected by the changes of prostate specific antigen (PSA) levels and maximum standardized uptake value (SUVmax) on 68Ga-PSMA-11 PET/CT imaging.Paired t test and Wilcoxon's sign rank test were used to analyze the data.Results No acute side effects were observed after therapy of 177Lu-PSMA-617.There were no statistically significant differences after therapy in WBC counts,RBC counts,and PLT,as well as Hb levels (t values:-0.28-1.11,all P＞ 0.05).No kidney toxicity was found.The PSA level after 177Lu-PSMA-617 therapy was significantly lower than that before therapy (80.70 (14.29,1 538.00) μg/L vs 604.60 (88.41,3 980.00) μg/L;u =59,P =0.023).Of the 11 patients,only 2 had elevated PSA levels and disease progression,while the other 9 patients had varying decreases,of which 2/11 decreased by ＞30％ and 7/11 decreased by ＞50％.After therapy,SUVmax of metastatic lesions and metastatic lymph nodes were decreased in 9 and 2 patients respectively.Conclusions 177Lu-PSMA-617 has a good therapeutic value for mCRPC.It is safe and has no obvious side effects.