Quantitative immunofluorescent estimation of estrogen receptor β expression in human solid tumors using flow cytometry

Богуш, Т. А.; Shaturova, A. S.; Dudko, E. A.; Zhuraev, E. E.; Полоцкий, Б. Е.; Ungiadze, G. V.; Давыдов, М. И.

doi:10.3103/s0027131411040031

Cited by 10 publications

(3 citation statements)

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“…The methodological approach used in this study, which was developed earlier to evaluate the expression of the multidrug resistance marker Pgp and estrogen receptor β [8,9], is devoid of subjectivity and allows strictly quantitative analysis of 5-10 thousand cells from a single cell suspension obtained from a suffi ciently large tumor specimen (up to 2 and even more cm in diameter). The vital importance of an integrated assessment of the level of expression of markers throughout the tumor site for accurate molecular diagnostics was convincingly demonstrated in a recent paper [11].…”

Section: Immunofluorescent Assay Of Ercc1 and Estimation Of Clinical mentioning

confidence: 99%

“…In this study, the parameters of ERCC1 expression in ovarian cancer tissue were estimated by immunoflu orescent analysis with the use of flow cytometry, which has been adapted for the study of tumor markers in solid tissues, is devoid of subjectivity, and allows a strictly quantitative analysis of 5-10 thousand cells from tumor specimens up to 2 cm in diameter and larger [8,9].…”

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confidence: 99%

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Immunofluorescent assay of ERCC1 and estimation of clinical significance of the protein expression in ovarian cancer tissue

Богуш

Dudko

Semakov

et al. 2014

Dokl Biochem Biophys

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Section: Immunofluorescent Assay Of Ercc1 and Estimation Of Clinical mentioning

confidence: 99%

mentioning

confidence: 99%

Immunofluorescent assay of ERCC1 and estimation of clinical significance of the protein expression in ovarian cancer tissue

Богуш

Dudko

Semakov

et al. 2014

Dokl Biochem Biophys

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“…Ранее мы адаптировали иммунофлуоресцентный метод, ассоциированный с проточной цитометрией, для изучения опухолевых маркёров в ткани сóлидных новообразований [4]. По сравнению с ИГХ-исследованием метод обладает рядом преимуществ: мягкая преаналитическая подготовка, строго количественная оценка результатов, нивелирование молекулярной гетерогенности опухоли за счет исследования маркёра в большом объеме опухолевого образца [2].…”

Section: Introductionunclassified

The INTRA-ASSAY PRECISION AND TIME STABILITY OF QUANTITATIVE ΒIII-Tubulin EXPRESSION ASSESSMENT IN SOLID TUMOR TISSUE

Башарина

Богуш

Рукавишникова

et al. 2020

Rossijskij bioterapevtičeskij žurnal

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Introduction. The introducing of tumor molecular profiling into clinical practice has revealed the need for development of new analytical methods for estimating marker expression in solid tumors, as routinely used method of immunohistochemistry has a number of significant drawbacks.Objective. Analytical validation of immunofluorescence staining and flow cytometry method developed by the authors for the examination of tumor protein markers in the solid tumors tissue.Materials and methods. Method validation was carried out by quantitative estimation of βIII-tubulin (TUBB3) expression in single-cell suspensions of non-small-cell lung cancer obtained from surgical tumor samples. Primary antibodies to TUBB3 (ab7751) and secondary DyLight 650-conjugated antibodies (ab98729) were used for immunofluorescent staining. The «Navios» flow cytometer (Beckman Coulter) was used to measure the fluorescence. The validation parameters were assessed by the coefficient of variation calculated as the ratio of standard deviation of TUBB3 level to its mean value.Results. Two parameters were analyzed: intra-assay precision and time stability of the results of the TUBB3 expression assessment. It was demonstrated that the mean coefficients of variation of the marker expression level in the tumor tissue did not exceed 20 % for both parameters. According to recommendations on the analytical validation of methods based on flow cytometry, it proves the validity of the method for these parameters.Conclusions. The intra-assay precision and time stability were demonstrated for the results of a quantitative estimation of TUBB3 expression in solid tumor tissue using immunofluorescence staining and flow cytometry method developed by the authors. The practical value of the time stability of immunofluorescence stain during 24 h storage of a stained cells suspension in the dark at 4 °C was highlighted. It shows the possibility of adjusting the time interval between completion of the analytical study part and flow cytometer measurement.

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The expression and clinical significance of ERβ/ERα in ovarian cancer: can we predict the effectiveness of platinum plus taxane therapy?

et al. 2021

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Quantitative immunofluorescent estimation of estrogen receptor β expression in human solid tumors using flow cytometry

Cited by 10 publications

References 7 publications

Immunofluorescent assay of ERCC1 and estimation of clinical significance of the protein expression in ovarian cancer tissue

Immunofluorescent assay of ERCC1 and estimation of clinical significance of the protein expression in ovarian cancer tissue

The INTRA-ASSAY PRECISION AND TIME STABILITY OF QUANTITATIVE ΒIII-Tubulin EXPRESSION ASSESSMENT IN SOLID TUMOR TISSUE

The expression and clinical significance of ERβ/ERα in ovarian cancer: can we predict the effectiveness of platinum plus taxane therapy?

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