Teresa A Almeida1, Antonieta Quispe-Ricalde2, Francisco Montes de Oca3, Pilar Foronda4, Mariano M Hernández5. 1. Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: tacosalm@ull.edu.es. 2. Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: maquispe@ull.es. 3. Hospital Quirón, Poeta Rodríguez Herrera 1, Santa Cruz de Tenerife, Tenerife 38006, Spain. Electronic address: fmontesdeoca@ginefem.com. 4. Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: pforonda@ull.es. 5. Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: mnhdez@ull.edu.es.
Abstract
OBJECTIVE: To evaluate 51 different housekeeping genes for their use as internal standards in myometrial and matched leiomyoma samples in proliferative and secretory phases. METHODS: RNA from 6 myometrium and matched leiomyoma samples was obtained from pre-menopausal women who underwent hysterectomy. Reverse-transcription and real-time quantitative PCR were achieved using TaqMan high-density open-array human endogenous control panel. RESULTS: Expression stability of 51 candidate genes was determined by GeNorm and NormFinder softwares. We identified 10 housekeeping genes, ARF1, MRPL19, FBXW2, PUM1, UBE2D2, EIF2B1, HPRT1, GUSB, ALAS1, and TRIM27, as the best set of normalization genes for comparing relative expression between leiomyoma and myometrium samples in proliferative and secretory phases. CONCLUSIONS: Adequate reference genes for accurate normalization are essential to compare gene expression between leiomyoma and myometrial samples. Ideal housekeeping genes must have stable expression patterns regardless of the sample type and menstrual cycle phase. In this study, we propose a set of 10 candidate genes with greater expression stability than those housekeeping genes commonly used in leiomyoma and myometrium tissues. Their use will improve the sensitivity and specificity of the gene expression analysis in these tissues.
OBJECTIVE: To evaluate 51 different housekeeping genes for their use as internal standards in myometrial and matched leiomyoma samples in proliferative and secretory phases. METHODS: RNA from 6 myometrium and matched leiomyoma samples was obtained from pre-menopausal women who underwent hysterectomy. Reverse-transcription and real-time quantitative PCR were achieved using TaqMan high-density open-array human endogenous control panel. RESULTS: Expression stability of 51 candidate genes was determined by GeNorm and NormFinder softwares. We identified 10 housekeeping genes, ARF1, MRPL19, FBXW2, PUM1, UBE2D2, EIF2B1, HPRT1, GUSB, ALAS1, and TRIM27, as the best set of normalization genes for comparing relative expression between leiomyoma and myometrium samples in proliferative and secretory phases. CONCLUSIONS: Adequate reference genes for accurate normalization are essential to compare gene expression between leiomyoma and myometrial samples. Ideal housekeeping genes must have stable expression patterns regardless of the sample type and menstrual cycle phase. In this study, we propose a set of 10 candidate genes with greater expression stability than those housekeeping genes commonly used in leiomyoma and myometrium tissues. Their use will improve the sensitivity and specificity of the gene expression analysis in these tissues.
Authors: P Asiabi; J Ambroise; C Giachini; M E Coccia; B Bearzatto; M C Chiti; M M Dolmans; C A Amorim Journal: J Assist Reprod Genet Date: 2020-07-30 Impact factor: 3.412
Authors: Magdalena Taube; Johanna C Andersson-Assarsson; Kristin Lindberg; Maria J Pereira; Markus Gäbel; Maria K Svensson; Jan W Eriksson; Per-Arne Svensson Journal: Adipocyte Date: 2015-05-28 Impact factor: 4.534
Authors: Febilla Fernando; Geertruda J M Veenboer; Martijn A Oudijk; Marlies A M Kampman; Karst Y Heida; Louise J M Lagendijk; Joris A M van der Post; Aldo Jongejan; Gijs B Afink; Carrie Ris-Stalpers Journal: Medicina (Kaunas) Date: 2021-05-21 Impact factor: 2.430