[Effects of artificial microRNA targeting glucosylceramide synthase on drug sensitivity of breast cancer cells].

Abstract

OBJECTIVE:

To explore the inhibitory effects on glucosylceramide synthase (GCS) expression and drug sensitivity in breast cancer cells by transfecting artificial microRNA targeting GCS.

METHODS:

Two microRNA expression vectors targeting GCS were constructed and transfected into MCF-7/ADR cells via Lipofectamine 2000. The levels of GCS mRNA and protein were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. Methyl thiazolyl tetrazolium (MTT) assay was used to assess the chemosensitivity of MCF-7/ADR cells to adriamycin (ADM) and vincristine.

RESULTS:

After transfection of two microRNA expression vectors, the expression of GCSmRNA in MCF-7/ADR cells was 0.098 ± 0.005 and 0.143 ± 0.007 respectively. Compared with the control cells (0.875 ± 0.008), the difference was significant (P < 0.01). The expression of GCS protein (0.127 ± 0.004, 0.165 ± 0.008) in MCF-7/ADR cells was lower than that in the control cells (0.765 ± 0.007; P < 0.01). Furthermore, in comparison with the control cells, the resistance factor to adriamycin significantly dropped to 4.06 and 6.06 while the drug resistance to vincristine decreased to 8.30 and 12.67 respectively (P < 0.01).

CONCLUSION:

Artificial microRNA targeting GCS inhibits the GCS expression and restores significantly the sensitivity of breast cancer cells to anticancer drugs. These findings may provide a novel strategy of enhancing the chemotherapy sensitivity of breast cancer.

• Authors: Yan-lin Sun, Peng Gao, Xiao-fang Zhang, Yong-sheng Gao, Geng-yin Zhou
• Journal: Zhonghua yi xue za zhi (Zhonghua Yi Xue Za Zhi) Vol. 92 Issue 46 Pg. 3296-9 (Dec 11 2012) ISSN: 0376-2491 [Print] China
• PMID: 23328518 (Publication Type: English Abstract, Journal Article, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• MicroRNAs
• RNA, Messenger
• Glucosyltransferases
• ceramide glucosyltransferase

Topics

• Breast Neoplasms (drug therapy, genetics)
• Drug Delivery Systems
• Drug Resistance, Neoplasm (drug effects)
• Female
• Glucosyltransferases (pharmacology, therapeutic use)
• Humans
• MCF-7 Cells
• MicroRNAs (genetics, therapeutic use)
• RNA, Messenger (genetics)