ABSTRACT
Objective:To investigate the effects of two SNP sites of delta-like ligand protein-1 (DLL1) gene rs2738822 (C>T) and rs9459988 (T>G) and gene expression on bone marrow suppression after neoadjuvant chemotherapy for breast cancer.Methods:Breast cancer patients who received neoadjuvant chemotherapy were selected as study subjects, including 90 patients with severe bone marrow suppression and 72 patients with mild bone marrow suppression. Patient’s demographic characteristics and laboratory test indicators were collected. Two SNP sites of DLL1, rs2738822 and rs9459988, were genotyped by capillary electrophoresis and section analysis (SNaPshot) . The relative mRNA expression of DLL1 gene was detected by quantitative reverse polymerase chain reaction (QRT-PCR) method.Results:For The rs2738822 of DLL1 gene, the genotype distribution difference between severe and mild bone marrow suppression groups was statistically significant ( χ2=8.622, P=0.013) . Compared with CC genotype, CT and TT genotype carriers had a higher risk of severe bone marrow suppression, with an OR value of 2.746 (1.335-6.882) and 3.054 (1.282-8.143) , respectively. The dominant model results showed that TT OR CT carriers had a significantly higher risk of severe bone marrow suppression than THOSE with CC genotype [ OR=2.976 (1.231-4.963) ]. For rs9459988, there was no significant difference in genotype distribution between severe bone marrow suppression group and mild bone marrow suppression group ( χ2=2.149, P=0.342) . Results of the dominant model showed that TG or GG carriers had a significantly higher risk of severe bone marrow suppression than TT carriers, with an OR value of 2.046 (1.053-5.611) . The relative mRNA expression level of DLL1 gene was 1.15±0.23 in patients with severe bone marrow suppression, which was significantly lower than that in patients with mild bone marrow suppression (2.64±0.51) ( t=6.381, P<0.001) . For rs2738822, with the increase of T allele, the relative mRNA expression level of DLL1 gene decreased gradually ( P<0.05) . For rs9459988, the relative mRNA expression level of DLL1 gene in patients with mutant allele G was also significantly lower than that in wild-type CC carriers ( P<0.05) . Conclusion:Mutations of DLL1 genes rs2738822 and rs9459988 are related to the occurrence of severe bone marrow suppression after neoadjuvant chemotherapy for breast cancer, and can be used as a genetic marker to predict the degree of bone marrow suppression after neoadjuvant chemotherapy for breast cancer patients.
ABSTRACT
Objective:To investigate the effect of c-maf gene on the MM cells' proliferation and invasion activity. Methods:Lipo-fectin Reagent was used to transfect c-maf siRNA into multiple myeloma cell of RPMI8226. The mRNA expression level of c-maf was detected by RT-PCR.Cell growth curve was measured by MTT assay. Transwell chamber test was used to measure MM cells' in vitro in-vasion activity. The cell cycle distribution were assessed by flow cytomentry. The protein expression levels of survivin,MMP-2, MMP-9, ARK5 and cyclin B1 were detected by Western blot. We also detected the activity of Caspase-3/7. Results:The c-maf siRNA was effectively transfected into cells and the mRNA expression of the c-maf gene was inhibited.MTT test and Transwell chamber test showed that the proliferation and in vitro invasion activity of transfected cells were significantly lower than those of other two groups (P<0.05). Cell cycle of c-maf siRNA transfected group cells was arrested in G2/M phase. The expression levels of survivin,MMP-2, MMP-9,ARK5,cyclin B1 and the activity of Caspase-3/7 between c-maf siRNA transfected group and the other two groups were sta-tistically different (P<0.05). Conclusion:c-maf gene by c-maf siRNA can remarkably inhibit proliferation and invasion of multiple my-eloma cell lines of RPMI8226. C-maf gene may be used as the target for multiple myeloma gene therapy.