Effects of MCCC2 knockdown on proliferation, migration and apoptosis of DU145 prostate cancer cells / 药学实践杂志

Objective To investigate the change of biological characteristics after stable knockdown of the coding gene of 3-methylcrotonyl-coenzyme A carboxylase β subunit (MCCC2) expression in DU145 by lentivirus shRNA. Methods Three groups were included in this study. shNC was the control group in which MCCC2 was negatively knocked down in DU145. shMCCC2 was the experimental group in which MCCC2 was knocked down. DU145 was the blank group without any treatment. The expression of MCCC2 was assessed by Western blot and qPCR. The proliferation of DU145 cells was detected by CCK8 assay. The migration ability of DU145 was detected by transwell. The apoptosis of DU145 cells was detected by flow cytometry. Results The expression level of MCCC2 in shMCCC2 group was significantly lower than that in shNC group (0.22 ± 0.02 vs 0.61 ± 0.06, P < 0.001). The proliferation (2.24 ± 0.04 vs 3.13 ± 0.15) and migration (23.96 ± 1.85 vs 49.73 ± 0.63) of DU145 cells in shMCCC2 group was significantly lower than that in shNC group, whereas the apoptosis (12.64 ± 0.30 vs 3.68 ± 0.02) of DU145 cells in shMCCC2 was significantly higher than that in shNC group. Conclusion MCCC2 knockdown significantly inhibited the proliferation and migration, and induced apoptosis of DU145 cells, which indicated that the down-regulation of MCCC2 is correlated with the change of tumor biological characteristics of DU145 cell line and can be a potential target for the treatment of prostate cancer.

Identification of a Novel Mutation in the MCCC2 Gene of a Korean Patient with 3-Methylcrotonyl-CoA Carboxylase Deficiency

3-methylcrotonyl-CoA carboxylase deficiency is an autosomal recessive disorder characterized by a defect in leucine catabolism. We report the case of an 80-day-old patient with 3-methylcrotonyl-CoA carboxylase deficiency who had elevated levels of 3-hydroxyisovalerylcarnitine (45.56 micromol/L; reference range, C (p.Gly105Arg)] at nucleotide position 313 and a mutation caused by a heterozygous A to T transversion [c.1252A>T (p.lle418Phe)] at nucleotide position 1252. Identification of these 2 novel MCCC2 gene mutations in our patient suggested that analysis of the MCCC1 and MCCC2 genes might prove useful in the diagnosis of 3-methylcrotonyl-CoA carboxylase deficiency.

A Case of Asymptomatic 3-methylcrotonylglycinuria Detected by Tandem Mass Spectrometry in Newborn Screening / 소아과

3-Methylcrotonylglycinuria is an autosomal recessive inborn error of leucine catabolism that results from the deficiency of 3-methylcrotonyl-CoA carboxylase(3-MCC). In 3-MCC deficiency 3-methylcrotonyl-CoA, may form glycine and carnitine conjugates. The primary metabolites are 3-hydroxyisovaleric acid(3-HIVA), 3-hydroxyisovaleryl carnitine(3-HIVC) and 3-methylcrotonylglycine(3-MCG). 3-hydroxyisovaleric acid(3-HIVA) and 3-methylcrotonylglycine(3-MCG) are increased in urine, and 3-hydroxyisovaleryl carnitine(3-HIVC) is found in blood and urine. 3-MCC is one of the four biotin- dependent carboxylases known in humans and is a heteromeric mitochondrial enzyme comprised of biotin-containing alpha-subunits and smaller beta-subunits. The gene for alpha-subunits(MCCC1) is located on chromosome 3q25-q27, beta-subunits(MCCC2) is located in 5q12-q13. Mutation in either of these genes may result in the deficiency of the enzyme activity. The introduction of tandem mass spectrometry in newborn screening has revealed an unexpectedly high incidence of this disorder and has also revealed that the range of clinical symptoms has become even wider ranging from neonatal onset with severe neurological involvement to asymptomatic newborns and adults. We report a case of a 43-day-old Korean asymptomatic girl with 3-Methylcrotonylglycinuria, detected by tandem mass spectrometry in newborn screening. This is resulted from the deficiency of 3-MCC by urine organic acid analysis. We found 2 mutations in the MCC2 gene of this patient. They are misssence mutation (D280Y) and splicing mutation(T357T). This patient on leucine restriction in conjunction with oral carnitine and glycine shows normal growth and development until now(10 months).