Abstract
AbstractThe aim of this study was to investigate the role of paclitaxel on RAB family of genes in primary breast cancer cell lines. The cancer breast cells obtained from 40 women during mastectomy were used to address this issue. The group included patients with intraductal breast cancer - lesions in I or II advancement level by TNM classification and G1-G2 by Bloom classification. (tumor dimensions up to 2.0 cm without metastases to lymph nodes). Cytostatic drugs before surgery were not administered to these patients. The cultures were conducted in 25 cm^2^ plastic containers at RPMI medium with addiction of 10% fetal bovine serum (FBS) at the standard conditions. After reaching concentration levels of 10 000/ml of the cells, the cultures were treated with 60 ng/ml and 300 ng/ml doses of paclitaxel. The concentrations were calculated in relation to therapeutic doses of paclitaxel, applied in polytherapy in patients with breast cancer. The cell cultures untreated for cytostatic were used as a control group. Analysis was conducted for RAB family of genes: RAB3D, RAB5B, RAB5C, RAB7, RAB7L1, RAB9P1, RAB10. RAB11A, RAB311B, RAB13, RAB18, RAB22A, RAB23, RAB26, RAB27A, RAB27B, RAB28, RAB30, RAB31, RAB33A, RAB3D6, RAB 38, RABL2B Total RNA was extracted from the harvest control group and the treated cells, and this was followed by cDNA synthesis, which was used for hybridization assays using arrays. A lower dose of paclitaxel (60 ng/ml) treatment resulted in an increase (2-4 fold- statistically significant), whereas a higher dose (300 ng/ml) caused a decrease (2-fold - statistically insignificant) in expression of examined oncogenes, compared to that of the control group.In summary, this data indicates that 60 ng/ml paclitaxel dose induced the RAB gene expression in an up-regulated pathway. A higher concentration of cytostatic (300 ng/ml) is a toxic dose for primary breast cells in vitro.
Highlights
In the metabolism of neoplasm cells, processes involving division and growth play an important role
It has been reported that alterations in gene expression involved in the cell cycle were characteristic of an increased activity of genes associated in cell proliferation and growth
BRCA1 belongs to the suppressor genes group, which is an activator of the transcription process, involved mainly in the DNA repairing system [2]
Summary
In the metabolism of neoplasm cells, processes involving division and growth play an important role. A significant number of these genes encode signaling proteins within the cell Some of these code factors influence cellular growth (sis, int) and growth factor receptors (erb-B, fms). In this group, there are known genes which code transcription factors (myb, myc, jun, fos), kinases (src,yes, abl, raf, mos), GTP binding proteins (ha-ras, ki-ras, n-ras) [1]. There are known genes which code transcription factors (myb, myc, jun, fos), kinases (src,yes, abl, raf, mos), GTP binding proteins (ha-ras, ki-ras, n-ras) [1] Some of these genes cause the transformation from normal cells to neoplastic cells when mutation in their structure occurs. The mentioned genes (RAB, ACT, MYC, JUN, FOS) can activate faster growth, division of cells and transformation processes
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