Assessing the Feasibility and Clinical Implications of multimarker RT-qPCR Profiling for Circulating Tumor Cell Detection in Breast Cancer Patients: A Pilot Study

Author(s) : Mehak Ali, Hafiz Muhammad Haseeb Khaliq, Amna Batool, Kausar Bano & Adnan Arshad

Abstract:

DOI: https://doi.org/10.52587/njmhs.v5i1.59

Objective:
We primarily aimed to detect, isolate and enumerate circulating tumor cells with the help of a panel of markers in a cohort of breast cancer patients in Pakistani Population. Based on clinical utility of CTCs we build a comparison between level of CTCs before and after at least one cycle of chemotherapy monitoring the therapeutic response by using CTCs as Prognostic marker.
Material and Methods:
EDTA Blood samples of Breast cancer female patients were collected at two different time’s points of their therapeutic regime. RNA was extracted and cDNA was synthesized. After isolation of CTCs, quantification of CTCs was determined with the help of a panel of markers (EpCAM, KRT & MAM) by using real time PCR during different time’s points of chemotherapy along with the formation of conclusive statistical relationships.
Results:
Our multi-marker analysis for CTCs on the basis of selected biomarkers is strongly associated with cancer manifestations including treatment efficacy and predicting its potential measures to metastasize. We found increase in expression of the three markers for CTCs in metastatic breast cancer patients as compared to the ones on neo adjuvant and adjuvant chemotherapeutic regimes. Moreover, the level of CTCs varied with molecular subtypes. HER2neu have more CTCs as compared to Luminal classified cancers.
Conclusion:
It will help the clinicians not only in determining personalized chemotherapeutic regimen but also help in stratifying patients into low and high risk of developing metastasis. It will further help to devise anti-metastatic therapies and improving prognosis. The prospect of CTCs is to be provided with the platform of analysis beyond just enumeration.

Keywords: Breast Cancer, CTCs, RT-qPCR, Liquid Biopsy, HER2neu