Ruthenium(II) complex containing cinnamic acid derivative inhibits cell cycle progression at G0/G1 and induces apoptosis in melanoma cells

Melanoma is a highly aggressive skin cancer with limited targeted therapy arsenal. The Ruthenium-based complexes have shown interesting pro-apoptotic effect on malignant tumor cell lines. In this study three Ruthenium(II)...

Berberine Inhibits MDA-MB-231 Cells by Attenuating Their Inflammatory Responses

Breast cancer initiation is closely associated with cytokines that can change the inflammatory tumor microenvironment. Compounds extracted from plants have been explored for the possibility of cancer treatment in the recent decades. Berberine is an isoquinoline plant alkaloid with remarkable antioxidant and anti-inflammation roles, which is used in ethnic medicines, including traditional Chinese and North American medicine. In the present study, we investigated the effects of berberine on the malignant tumor cell behaviors in a breast cell line, MDA-MB-231. We found that berberine could not influence the cell viability in normal condition but was able to decrease the cancer cell migration capability in a scratch wound model and accordingly prolong the wound healing time. Furthermore, our results demonstrated that berberine inhibited the increased phosphorylation of c-Jun and c-Fos in these scratched cancer cells. With the cotreatment with LPS, which could boost the expression of cytokines in these cancer cells, berberine significantly reduced the increased expression of TNF-α and IL-6. Meanwhile, we found that berberine inhibited the activation of NF-κB by preventing the degradation of IκBα.

Molecular Cloning, Expression and Purification of G-CSF Isoform D, an Alternative Splice Variant of Human G-CSF

Granulocyte colony-stimulating factor (G-CSF) is the major regulator of hemopoiesis and granulopoiesis. However, overexpression of G-CSF has been implicated in several important processes in tumor biology such as tumor growth, angiogenesis, and metastasis. Four different mRNA isoforms resulting from alternative splicing have been reported for G-CSF (transcript variants 1, 2, 3 and 4). The mRNAs and protein products of splice variants 1 and 2 have been isolated for the first time, from tumor cell lines. In the present study for the first time we isolated the G-CSF transcript variant 4 encoding G-CSF isoform D from a highly malignant tumor cell line (Mehr80) with overexpression of G-CSF. Both the full-length G-CSF isoform B and G-CSF isoform D were cloned from Mehr80 cell line, overexpressed in Escherichia coli as N-terminal glutathione-S-transferase fusion proteins in the form of inclusion bodies and affinity purified by the batch method using glutathione-Sepharose 4B resin. Both fusion proteins were successfully cloned and expressed. Folded recombinant proteins were solubilized from inclusion bodies using sarkosyl, Triton X-114 and CHAPS and purified. The purity of G-CSF isoforms was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and they were clearly detected in western blot analysis using anti-G-CSF polyclonal antibody. The G-CSF plays various roles in physiological and pathological conditions, however to date, the differential function of G-CSF isoforms remains unknown. Considering the fact that G-CSF isoform D was isolated from a highly malignant tumor cell line with overexpression of G-CSF, the role of this splice variant in tumorigenesis requires further investigation.

Detection of SNPs of MTHFR Genes in Five Hematological Malignant Tumor Cell Lines

Abstract 5130 Background & Aim: This study detected three single nucleotide polymorphisms (SNPs) of methylenetetrahydrofolate reductase (MTHFR) gene in five hematology malignant cell lines, to find correlations between MTHFR gene SNPs and hematological malignancies by horizontal comparison. Materials & Methods: Cell line K562, Ka, HL-60, U937, and Raji were cultured. The genomic DNA was isolated by QIAamp DNA Blood Mini kit. Designing primes were amplified by PCR to get the related DNA fragments. MTHFR gene A1298C (rs1801131), C677T (rs1801131), G1793A (rs2274976) was genotyped by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method (MALDI-TOFMS). Results: The results showed that the genotype of locus A1298C (rs1801131) on MTHFR gene was AA for U937 cell line. While for K562, Ka, HL-60, and Raji cell lines, that was CA. For all the five hematology malignancies cell lines, the genotype of locus C677T (rs1801133) on the MTHFR gene was CC, and that of locus G1793A (rs2274976) was GG. Conclusion: The genotype of locus A1298C (rs1801131) on MTHFR gene was not all the same in the detected hematology malignant cell lines, while the genotypes of the other 2 loci were same in the 5 cell lines. Disclosures: No relevant conflicts of interest to declare.

Detection of SNPs of Dck and Cda Genes in Five Hematological Malignant Tumor Cell Lines

Abstract 5129 Background & Aim: This study detected single nucleotide polymorphisms (SNPs) of deoxycytidine kinase(DCK) gene, cytidine deaminase(CDA) gene in five hematological malignant tumor cell lines, to find connections between sNPs and Cytarabine dose selection in the treatment of some hematopathy. Materials & Methods: Cell line K562, Ka, HL-60, U937, and Raji were cultured. The genomic DNA was isolated by QIAamp DNA Blood Mini kit. Designing primes were amplified by PCR to get the related DNA fragments. DCK gene A674G(rs111454937) C1644T(rs72552079), CDA gene A79C(rs2072671) G208A(rs60369023) was genotyped by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method (MALDI-TOF MS). Results: The genotype of locus A674G(rs111454937) on DCK gene in all five hematology system cell lines was AA; The genotype of locus C1644T(rs72552079) on DCK gene in all five hematology system cell lines was CC. The genotype of locus G208A(rs60369023) on CDA gene in all five hematology system cell lines was GG; For HL-60, U937, and Raji cell lines, the genotype of locus A79C(rs2072671) on CDA gene was AA, while for K562 and Ka cell lines, that was C/A. Conclusions: The genotype of locus A79C(rs2072671) on CDA gene was not all the same in the detected hematological malignant tumor cell lines, while the genotypes of the other 3 loci were same in the 5 cell lines. Disclosures: No relevant conflicts of interest to declare.