Manjula Y Rao

Publications (3)7.74 Total impact

• Chapter: Purification of Diseased Cells from Barrett’s Esophagus and Related Lesions by Laser Capture Microdissection

  Masood A. Shammas, Manjula Y. Rao
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  ABSTRACT: Barrett’s esophageal adenocarcinoma (BEAC) arises from Barrett’s esophagus (BE), a premalignant lesion caused by acid reflux (heartburn). Although the cancer is uncommon, its incidence is rapidly rising in western countries. Like most other cancers, BEAC cells also have elevated telomerase activity which maintains telomere length and supports continued proliferation of these cells. It is not clear if telomerase is activated early at premalignant (BE) stage, because reports of telomerase activity in Barrett’s and normal esophagi have been controversial. We have shown that detection of telomerase and telomeres becomes easier and much more reliable if purified BE cells are used instead of tissue specimens. This chapter, therefore, emphasizes the importance of laser capture microdissection and provides the method to purify Barrett’s esophagus related cells, using this technique. Key wordsLaser microdissection–Barrett’s esophagus–Esophageal adenocarcinoma–Esophageal metaplasia–Telomere–Telomerase
  07/2011: pages 181-187;
• Article: Purification of diseased cells from Barrett's esophagus and related lesions by laser capture microdissection.

  Masood A Shammas, Manjula Y Rao
  [show abstract] [hide abstract]
  ABSTRACT: Barrett's esophageal adenocarcinoma (BEAC) arises from Barrett's esophagus (BE), a premalignant lesion caused by acid reflux (heartburn). Although the cancer is uncommon, its incidence is rapidly rising in western countries. Like most other cancers, BEAC cells also have elevated telomerase activity which maintains telomere length and supports continued proliferation of these cells. It is not clear if telomerase is activated early at premalignant (BE) stage, because reports of telomerase activity in Barrett's and normal esophagi have been controversial. We have shown that detection of telomerase and telomeres becomes easier and much more reliable if purified BE cells are used instead of tissue specimens. This chapter, therefore, emphasizes the importance of laser capture microdissection and provides the method to purify Barrett's esophagus related cells, using this technique.
  Methods in molecular biology (Clifton, N.J.) 01/2011; 755:181-7.
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  Available from: rajgoyal.com

  Article: Telomere maintenance in laser capture microdissection-purified Barrett's adenocarcinoma cells and effect of telomerase inhibition in vivo.

  Masood A Shammas, Aamer Qazi, Ramesh B Batchu, Robert C Bertheau, Jason Y Y Wong, Manjula Y Rao, Madhu Prasad, Diptiman Chanda, Selvarangan Ponnazhagan, Kenneth C Anderson, Christopher P Steffes, Nikhil C Munshi, Immaculata De Vivo, David G Beer, Sergei Gryaznov, Donald W Weaver, Raj K Goyal
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  ABSTRACT: The aims of this study were to investigate telomere function in normal and Barrett's esophageal adenocarcinoma (BEAC) cells purified by laser capture microdissection and to evaluate the effect of telomerase inhibition in cancer cells in vitro and in vivo. Epithelial cells were purified from surgically resected esophagi. Telomerase activity was measured by modified telomeric repeat amplification protocol and telomere length was determined by real-time PCR assay. To evaluate the effect of telomerase inhibition, adenocarcinoma cell lines were continuously treated with a specific telomerase inhibitor (GRN163L) and live cell number was determined weekly. Apoptosis was evaluated by Annexin labeling and senescence by beta-galactosidase staining. For in vivo studies, severe combined immunodeficient mice were s.c. inoculated with adenocarcinoma cells and following appearance of palpable tumors, injected i.p. with saline or GRN163L. Telomerase activity was significantly elevated whereas telomeres were shorter in BEAC cells relative to normal esophageal epithelial cells. The treatment of adenocarcinoma cells with telomerase inhibitor, GRN163L, led to loss of telomerase activity, reduction in telomere length, and growth arrest through induction of both the senescence and apoptosis. GRN163L-induced cell death could also be expedited by addition of the chemotherapeutic agents doxorubicin and ritonavir. Finally, the treatment with GRN163L led to a significant reduction in tumor volume in a subcutaneous tumor model. We show that telomerase activity is significantly elevated whereas telomeres are shorter in BEAC and suppression of telomerase inhibits proliferation of adenocarcinoma cells both in vitro and in vivo.
  Clinical Cancer Research 09/2008; 14(15):4971-80. · 7.74 Impact Factor