ABSTRACT
OBJECTIVE@#To investigate the expression of C/EBPα gene in elderly patients with multiple myeloma （MM） and its prognostic significance.@*METHODS@#Sixty-nine olderly patients with multiple myeloma (MM) treated in our hospital from February 2015 to October 2017 were selected and enrolled in the MM group, 38 healthy persons received physical examination were selected and enrolled in the control group. The bone marrow of 2 groups was collected and the mononuclear cells were isolated.The mRNA expression level of C/EBPα gene in mononuclear cells was determined by RT-PCR, the Western blot was used to detect the protin expression level of PBMNC C/EBPα, and the protein level of C/EBPα in bone marrow was detected by immunohistochemistry. The correlations of C/EBPα gene expression with the clinical characteristics and survival time in MM patients were analyzed.@*RESULTS@#The expression level of mRNA and protein of C/EBPα in MM patients was significantly lower than that in the control group (P0.05).Immunohistochemical staining showed that the bone marrow samples of the control group were stained more deeply, and the staining intensity in bone marrow samples of MM patients with CR, PR and relapse was successively descended. The protein level of C/EBPα in CR patients with MM was significantly higher than that in PR and relapsed patients by Western blot (P<0.05). Kaplan-Meier survival analysis showed that OS and DFS in the patients with high expression of C/EBPα gene were higher than those in low expression group (P<0.05). Multivariate Cox regression analysis showed that CRP,ratio of myeloma cells and C/EBPα gene were independent factors affecting OS and PFS (P<0.05).@*CONCLUSION@#The expression level of C/EBPα gene in MM patients is low that may stimulate the genesis of MM, and the expression of C/EBPα gene closely relates with the development of MM disease.
Subject(s)
Aged , Humans , Bone Marrow , CCAAT-Enhancer-Binding Protein-alpha , Multiple Myeloma , Genetics , Neoplasm Recurrence, Local , PrognosisABSTRACT
Objective To investigate the correlation between the expression of aryl hydrocarbon receptor(AHR) mR-NA and tryptophan dioxygenase (TDO) mRNA in bone marrow mononuclear cells of patients with acute leukemia.Methods Sixty-five patients with newly diagnosed acute leukemia in Henan Provincial People's Hospital from August 2013 to August 2014 were selected as observation group,and there were 50 patients with acute myeloid leukaemia(AML) and 15 patients with acute lymphoblastic leukaemia(ALL).Fifteen patients with anemia were selected as control group in the same period(excluding the malignant disease of blood system).The expression of AHR mRNA and TDO mRNA in bone marrow mononuclear cells of patients in the groups was detected by real time fluorescence quantitative reverse transcription polymerase chain reaction.The correlation between AHR mRNA and TDO mRNA was analyzed.Results The expression of TDO mRNA and AHR mRNA in bone marrow mononuclear cells of AML and ALL patients in the observation group was significantly higher than that in the control group(P ＜0.05).There was no significant difference in the expression of TDO mRNA and AHR mRNA in bone marrow mononuclear cells between AML and ALL patients (P ＜ 0.05).There was significantly positive correlation between the expression of TDO mRNA and AHR mRNA in bone marrow mononuclear cells of AML and ALL patients(r =0.801,0.922;P ＜ 0.05).The levels of white blood cell,hemoglobin,platelet and lactate dehydrogenase were not related to the expression of TDO mRNA and AHR mRNA in AML and ALL patients(P ＜ 0.05).Conclusion The expression of TDO and AHR in bone marrow mononuclear cells of acute leukemia patients is high,and the TDO-KYN-AHR pathway promotes the development of acute leukemia.
ABSTRACT
Objective To establish a method for specific gene editing of the second exon of mouse MAD2L1 gene by CRISPR/Cas9, and analyze its off-target effect. Methods The gene editing site for MAD2L1 gene was designed by CHOP?CHOP, and the Cas9?MAD2L1 vector was constructed based on the designed editing site. Cas9?MAD2L1 was then transfected into NIH/3T3 cells and screened with puromycin, followed by observing GFP expression using fluorescence microscopy. The genomic DNA from transfected cells was extracted and a partial fragment of MAD2L1 gene was amplified by PCR. T7E1 analy?sis and Sangger sequencing were used for gene editing and off?target analysis. Results After Cas9?MAD2L1 transfection and puromycin screening, a large number of GFP?expressing cells were observed under the fluorescence microscope. Combined the PCR result with TE71 analysis, the amplified 228 bp PCR products can be digested into 166 bp and 62 bp fragments. The se?quencing result showed that the second exon of MAD2L1 gene was successfully edited, and the off?target effect was undetected in our system. Conclusions The method for specific gene editing of the second exon of mouse MAD2L1 gene by CRISPR/Cas9 is successfully established, and off?target effect of MAD2L1 gene is not detected.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the expressions of metastasis-associated protein (S100A4) and matrix metalloproteinase 9 (MMP9) in human non-small cell lung cancer (NSCLC) and investigate their correlations to the infiltration, metastasis and prognosis of NSCLC.</p><p><b>METHODS</b>The expressions of S100A4 and MMP9 were detected in 41 NSCLC specimens and 6 normal lung tissue specimens using immunohistochemistry with SP method. Univariate and multivariate survival analysis were used to analyze the correlations of S100A4 and MMP9 to the clinicopathological characteristics and progrnosis of NSCLC.</p><p><b>RESULTS</b>Compared with normal lung tissues, NSCLC showed significantly increased positivity for S100A4 and MMP9 expression (P<0.05); their expression were significantly higher in adenocarcinoma than in squamous cell carcinoma (P<0.01), and higher in metastatic NSCLC than in that without lymphatic metastasis (P<0.01). The positive expression rates of S100A4 and MMP9 were significantly higher in tumors in TNM stages III +IV than in stages II+I (P<0.05). S100A4 expression was positively correlated to tumor size (P<0.001), while MMP9 was inversely correlated to tumor differentiation (P<0.05). The expressions of S100A4 and MMP9 were both correlated to lymphatic metastasis, TNM stages and pathological types (P<0.05), and they also showed a mutual correlation (P<0.01). Univariate survival analysis confirmed the effects of histological types, lymphatic metastasis, clinical TNM stages and expressions of S100A4 and MMP9 on the survival time of NSCLC patients (P<0.001). Multivariate survival analysis identified clinical TNM stages and expressions of S100A4 and MMP9 as the independent factors affecting the prognosis of NSCLC (P<0.05).</p><p><b>CONCLUSION</b>The expressions of S100A4 and MMP9 are up-regulated in NSCLC and have significant correlations to the clinical and biological behaviors of NSCLC. S100A4 and MMP9 status are independent prognostic predictors of NSCLC, and detection of their expressions may help evaluate the prognosis of NSCLC.</p>