A Prospective Study of Validation of MicroRNAs to Assess Response and Prognosis among Cervical Cancer Treated with Radical Image-guided Chemoradiotherapy- MARC Study

Cervical cancer is a major cause of mortality worldwide as well as in India. The standard treatment for locally advanced cervical cancer (LACC) patients consists of radiotherapy (45 Gray in 25 Fractions over 5 weeks) in combination with weekly cisplatin/carboplatin-based chemotherapy. Unfortunately, nearly 20-30% of patients with cervical cancer have an unfavourable response to conventional chemoradiation treatment; these patients have a higher recurrence rate and worse survival in the first five years. We need to identify certain biomarkers to predict the clinical response in cervical cancer patients receiving chemoradiation and correlate with prognosis. MicroRNAs(miRNAs) are potential biomarkers in cervical cancer; however, their role in identifying patients who do not respond to conventional treatment remains poorly investigated. This study is aimed to evaluate some potential miRNAs which have shown good correlation in invitro to be evaluated in clinical settings and correlate with response and prognosis.

40 patients with biopsy proven cervical cancer will be recruited for the study. Informed consent will be taken prior to enrollment in the study. Standard treatment of cervical cancer will be followed. Tissue biopsy/brush sampling will be obtained at baseline prior to treatment and at 3 months after chemoradiation. The blood samples will be collected at the following time points – 1. Diagnosis 2. Completion of EBRT 3. Completion of Brachytherapy 4. 3 months after the completion of treatment 5. 1 year after treatment 6. 2 years after treatment 7. At recurrence or metastases.

We plan to generate patient specific cell lines which will be used for validating the clinical endpoints. The tumor biopsies taken from the patients will be washed with phosphate buffered saline, will be incubated with antibiotic–antimycotic for 20 s, washed once in PBS, incubated in gentamicin for 20 s and then washed in PBS. The tissue will be cut into small pieces, treated enzymatically with collagenase for 1 hrs at 37 ºC and placed in a cell culture dish for 10 min to attach and cultured in the presence of Epithelial Cell Growth Medium containing 10% FBS. The cells will be harvested by trypsinization, and western blotting will be carried out for pan-cytokeratin, cytokeratin 8 and E-cadherin to confirm the cells. These cell lines will be used to validate the findings obtained patient data. Response assessment and follow up schedule: After completion of chemoradiation and brachytherapy, patient will be assessed at 1 month post radiation to assess response and toxicity. The response at 3 months will be assessed clinically and with MRI pelvis as per routine practice. The follow up schedule is 3 monthly till two years and then 6 monthly till 5 years. Thereafter patients are followed up yearly.