Abnormal Expression of C3ORF9 Gene in Patients with Myelodysplastic Syndromes

Abstract

Myelodysplastic Syndrome (MDS) cells present genetic instability and dysregulation of the gene C30RF9, which was isolated from an MDS cDNA library and codes for a putative protein. We studied the expression of C30RF9 in MDS syndromes to contribute to the understanding of the pathophysiology of MDS. One hundred and thirty-one patients and 35 healthy controls were involved in our study. Bone marrow aspirates and isolated CD34+ cells were used. Gene expression was estimated by quantitative PCR. C3ORF9 was found to be down-regulated in patients with CMML compared to the controls (p<0.01). There was no difference between RARS and the controls (p=0.1), while increased expression was found in RA, RAEB and RAEB-T (p< 0.01 for all). No mutations or polymorphism were detected in our population. CD34+ cells expressed higher levels of C30RF9 (p<0.01) in patients. The gene expression was correlated to the percentage of + cells in RAEB and RAEB-T (r= 0.64). The altered C30RF9 expression was possibly due to different gene regulation in these patients and/or to the increased CD34+ cells.

Myostatin Expression in MDS Patients with Wasting Syndrome

“…In detail, 500 mg of muscle was homogenized in 3.75 mL TRIZOL LS (Life Technologies) with a tissue homogenizer. RNA was extracted from the homogenate by adding 1 mL chloroform, incubating at 25EC for 15 min, centrifugation at 12000 g for 15 min and collecting the upper aqueous phase 19 . RNA was precipitated using isopropyl alcohol according to standard protocol 19,[21][22][23] .…”

“…The RT-PCR was performed using 1 µg of RNA from patients and healthy individuals using the SuperScript TM One-Step RT-PCR kit (Invitrogen, Carlsbad, USA) as described earlier 19 .…”

“…The β-actin amplification was done as described elsewhere and a product of 290 bp in the 3'-UTR of the mRNA was obtained 19 .…”

“…A final extension cycle was made 10 min at 72EC. The DNA Engine Peltier Gradient Thermal Cycler (Bio-Rad Ltd. Corston Bath UK) was used 19 .…”

“…We examined the expression of the GDF-8 gene in patients with MDS to study mechanisms involved in the development of the cachexia syndrome in these patients 19 . We also searched for possible agents influencing myostatin expression in these patients.…”