Wannume Henry, Niyonzima Nixon, Kalungi Sam, Okuni Julius Boniface, Okecha Tonny, Kakungulu Edward, Kiwuwa Steven Mpungu, Waiswa Geoffrey, Kadhumbula Sylvester, Namayanja Monica, Nabwana Martin, Orem Jackson
Department of Clinical Support Services, Division of Laboratory and Pathology Medicine, Uganda Cancer Institute, Kampala, Uganda.
Department of Biomolecular Resources and Biolab Sciences, College of Veterinary Medicine, Animal Resources and Biosecurity, Makerere University, Kampala, Uganda.
PLoS One. 2025 Jan 3;20(1):e0311185. doi: 10.1371/journal.pone.0311185. eCollection 2025.
The detection of Estrogen Receptor (ER), Progesterone Receptor (PR), and Human epidermal growth factor receptor 2 (HER-2) is important for the stratification of breast cancer and the selection of therapeutic modalities. This study aimed to determine the quantitative expression of ER, PR and HER-2 using Immunohistochemistry and their correlation with quantitative baseline Ct values measured using Quantitative Polymerase Chain Reaction (PCR). This study also assessed the use of fresh breast tissue biopsies preserved in RNAlater solution in the quantitative detection of these receptors using PCR technique. The study evaluated 20 matched formalin fixed paraffin embedded and RNAlater preserved samples for ER, PR, and HER-2 using IHC and quantitative PCR technique. One portion of the breast tissue biopsy was fixed immediately in 10% neutral buffered formalin and another was preserved in RNAlater. After the histological confirmation of breast cancer by the H&E technique, formalin fixed paraffin embedded tissues (FFPE)-positive cases were matched with their corresponding RNAlater samples for IHC and qPCR. The extracted RNA was quantified using Nanodrop technology, resulting into complementary DNA. ER and PR using IHC were expressed in 60% (n = 12) of the study samples and were negative in 40% (n = 8) of samples. HER-2 was negative in 70% (n = 14) of study samples, 25% (n = 5) positive, and 5% (n = 1) equivocal. With the quantitative expression of ER, PR, and HER-2 being reported in the IHC triple-negative breast cancer cases. The mean Ct values for the hormonal receptors correlated with what has been previously studied with ER at 19.631, PR at 25.410 and HER-2 at 25.695. There was no statistically significant difference between the mean Ct values of RNAlater and FFPE with their P-values being 0.9919, 0.0896 and < 0.0001 for ER, PR, and HER-2 respectively. P-values; 0.9919 and 0.0896 for ER and PR respectively being greater than 0.05 it's a borderline significance although HER-2 had a statistical significance. With a concordance in the detection of these breast cancer hormonal receptors, qPCR can be used in our setting considering the delays that may be associated in following the samples through IHC processing.
雌激素受体(ER)、孕激素受体(PR)和人表皮生长因子受体2(HER-2)的检测对于乳腺癌的分层及治疗方式的选择至关重要。本研究旨在通过免疫组织化学法测定ER、PR和HER-2的定量表达,并研究其与采用定量聚合酶链反应(PCR)测得的基线定量Ct值之间的相关性。本研究还评估了保存在RNA Later溶液中的新鲜乳腺组织活检样本在使用PCR技术定量检测这些受体中的应用。该研究使用免疫组织化学(IHC)和定量PCR技术对20对匹配的福尔马林固定石蜡包埋样本及保存在RNA Later中的样本进行了ER、PR和HER-2评估。乳腺组织活检样本的一部分立即固定于10%中性缓冲福尔马林中,另一部分保存在RNA Later中。在通过苏木精-伊红(H&E)技术进行乳腺癌组织学确认后,福尔马林固定石蜡包埋组织(FFPE)阳性病例与其对应的RNA Later样本进行IHC和qPCR检测。提取的RNA使用Nanodrop技术进行定量,然后生成互补DNA。在研究样本中,通过IHC检测,ER和PR在60%(n = 12)的样本中呈阳性表达,在40%(n = 8)的样本中呈阴性。HER-2在70%(n = 14)的研究样本中呈阴性,25%(n = 5)呈阳性,5%(n = 1)为可疑阳性。在免疫组织化学检测的三阴性乳腺癌病例中报告了ER、PR和HER-2的定量表达。激素受体的平均Ct值与之前对ER的研究结果相关,ER为19.631,PR为25.410,HER-2为25.695。RNA Later样本和FFPE样本的平均Ct值之间无统计学显著差异,ER、PR和HER-2的P值分别为0.9919、0.0896和<0.0001。ER和PR的P值分别为0.9919和0.0896,大于0.05,虽为临界显著性,但HER-2具有统计学显著性。鉴于这些乳腺癌激素受体检测结果具有一致性,考虑到通过免疫组织化学处理样本可能会出现延迟,在我们的研究环境中可使用定量聚合酶链反应。