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Objective To investigate the correlation between the expression level of BLM and its clinical significance in leukemia.Methods 125 bone marrow specimens of inpatients and outpatients with leukemia were collected in Anhui provincial hospital from January 2011 to December 2011.125 leukemia patients were diagnosed and classified into acute leukemia (AL,n =66) and chronic myelogenous leukemia (CML,n =59) by Morphologic and Immunologic criteria,5 non-tumor individuals were included as control group.The BLM mRNA expressions were by reverse transcription-polymerase chain reaction(RT-PCR).The specimens were devided into groups according to the age,gender,leukemia type,peripheral blood leukocyte counts,hepatomegalia and(or) splenomegaly,fusion gene,chromosome karyotype,whether first visit and transplantation.The expression of BLM gene in each group and the correlation with above factors were retrospectively analysed.The statistical methods such as chi-square test,single factor variance analysis,t test and Pearson correlation test were mainly used.Results BLM mRNA was detected in leukemia.In bone marrow cells,the BLM gene expression was positive in 71 patients and negative in 54 patients.But none of 5 non-tumor bone marrow cells expressed BLM gene.The difference of BLM expression between patients and controls was statistically significant in two groups,i.e.peripheral blood leukocyte counts and fusion gene (x2 =14.730,22.399 ; P ＜ 0.05),but there is no statistical significant differences in other groups.The expressions of BLM mRNA in leukemia patients who had been treated with chemotherapy were lower than those newly diagnosed (0.1788 ± 0.1091 vs 0.3276 ± 0.2016 ; P ＜ 0.05).Moreover,BLM mRNA level in post-bone marrow transplant patients was lower than those not treated (0.1271 ± 0.1009 vs 0.2902 ±0.2034 ; P ＜ 0.05).Pearson correlation analysis showed that higher BLM mRNA expression positively correlated with fusion gene (r =0.357,P ＜ 0.01) and chromosome abnormality (r =0.279,P ＜ 0.05).Conclusion The BLM mRNA expression level of measurement can be used as judgment for leukemia patients disease severity and the index of prognosis,testing the level may provide a basis for clinical and curative effect judgment.
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Objective To establish a real-time fluorescence quantitative PCR method for detection of the different expression level of FPGS in methotrexate enantiomer-resistant A549 cell lines,and observe FPGS mRNA expression in patients with leukemia.Methods A real-time fluorescence quantitative PCR method for detection FPGS mRNA was established using SYBR Green Ⅰ as fluorescence and β-actin as reference.The method was evaluated by Ct,correlation coefficient,slope,repeatability curve,melting curve and amplification efficiency curve.The expression levels of FPGS gene in methotrexate enantiomer-resistant A549 cell lines and methotrexate resistant leukemia cells in bone marrow were detected by the method.Results The standard curves had a high linear relationship between cycle threshold and template concentration.The correlation coefficients of FPGS and β-actin were 0.996 8 and 0.998 7,and the slopes were -3.595 and -3.740,respectively.The inter-coefficient of variation was from 1.27% to 2.95%.The intra-coefficient of variation was 3.82%.The method was characterized with specific melting curve and similar amplification efficiency(slope was 0.021 7).The relative contents of FPGS mRNA were(3.51 ±0.66),(0.16 ±0.01) and(1.00 ±0.31) in L-(+)-MTX/A549 cells(L),D-(-)-MTX/A549 cells(D)and A549 parent cells,and there was statistically difference among the three groups(F = 64.45 ,P＜ 0.01)Statistical difference was observed between L and D(q =9.29,P＜0.01).After treated with MTX,the expression level of FPGS mRNA was(0.35 ± 0.04) in methotrexate resistant leukemia patients,compared with(1.00 ± 0.44) before treatment.Statistical difference was observed(t = 8.83 ,P＜ 0.01).Conclusions The real-time fluorescence quantitative PCR is suitable for the quantification of FPGS.The expression levels of FPGS in methotrexate resistant leukemia cells in bone marrow and drug resistant cells are different.Two enantiomer forms of methotrexate may play different roles in drug resistance mechanisms.
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<p><b>OBJECTIVE</b>To study the antitumor activities of the triterpene glycoside colochiroside A (CA) from the sea cucumber Colochirus anceps.</p><p><b>METHOD</b>The tests of antitumor activities in vitro and in vivo were applied to demonstrate the effect of CA.</p><p><b>RESULT</b>The preliminary cytotoxic assay of CA exhibited significant cytotoxic activity against 6 types cultured tumor cell lines of p388, HL60, A-549, SpC-A4, MKN-28, and SGC-7901, the mean of IC50 were (3.61 +/- 0.55) mg x L(-1). The preliminary antitumor assay of CA indicated that this saponin exhibited high inhibiting activity against the H22 live cancer and the S180 sarcoma cells in mouse. The inhibition ratio to H22 liver cancer were 34.8%, 43.9% and 52.2%, while the ratio to S180 sarcoma were 36.4%, 70.0%, the immunoregulatory founction study indicated CA has not significant effect on the developments of thymus and spleen.</p><p><b>CONCLUSION</b>The saponin CA exhibited remarkable antineoplastic activities in vitro and in vivo, and could not reduce the immunoregulatory founction of mice.</p>
Subject(s)
Animals , Mice , Antineoplastic Agents , Pharmacology , Cell Line, Tumor , Glycosides , Pharmacology , Neoplasms, Experimental , Drug Therapy , Sea Cucumbers , Chemistry , Triterpenes , PharmacologyABSTRACT
Objective To search for triterpene glycoside with new structure. Methods We studied the sea cucumber Colochirous anceps belonging to the family cucumariidea with many chromatography methods and found a new sulfated triterpene glycoside, which structure was established by a combination of spectroscopic analysis (IR, UV, ESI-MS and 2D-NMR), chemical transformations and other chemical evidence. Results and Conclusion The new sulfated triterpene glycoside was isolated and named Colochiroside A (1) .
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Objective To study the active compounds of the sea cucumber Colochirus anceps.Methods The compounds were isolated and purified from the low-polarity part of EtOH extract of the sea cucumber,and their structures were established by a combination of spectroscopic analysis,chemical transformation,and other chemical evidence.Results Four compounds were isolated and their structures were established as 1-O-?-D-glucopyranosyl-(2S,3R,4E,8E)-2-docosanoylamino-13-methyl-4,8-hexadecadiene-1,3-diol(Ⅰ),1-O-?-D-glucopyranosyl-(2S,3R,4E,8E)-2-[(2R)-2-hydroxydocosa-noylamino]-13-methyl-4,8-hexadecadiene-1,3-diol(Ⅱ),1-O-?-D-glucopyranosyl-(2S,3R,4E,8E)-2-[(2R)-2-hydroxy-tricosanoylamido]-13methyl-4,8-hexadecadiene-1,3-diol(Ⅲ),and 1-O-?D-glucopyranosyl-(2S,3R,4E,8E)-2-[(2R)-2-hydroxy-tetracosanoylamino]-13-methyl-4,8-hexadecadiene-1,3-diol(Ⅳ).Conclusion Four new cerebrosides are isolated from the sea cucumber C.anceps for the first time.