Next-generation sequencing unravels extensive genetic alteration in recurrent ovarian cancer and unique genetic changes in drug-resistant recurrent ovarian cancer

Abstract

Background: By using a high-throughput sequencing technique, we sought to delineate genetic alterations in recurrent ovarian cancer patients and further compare genetic changes in drug-resistant and -sensitive recurrent ovarian cancer patients. We also sought to study the specificity, sensitivity, and consistency of DNA biomarkers in liquid biopsy specimens and ovarian cancer tissue DNA. Methods: Tumor tissue specimens and blood samples were obtained from pathologically proven recurrent ovarian cancer patients. Genomic DNA was extracted from tumor tissues, blood cells, ascites, and urine samples. The DNA Library was constructed and sequencing was performed using the Illumina HiSeq 4000 high-throughput sequencing platform. Bioinformatic analysis was done using the Torrent Suite software. Results: Ten patients with pathologically proven drug-resistant recurrent ovarian cancer and 11 patients with sensitive recurrent ovarian cancer were included. The 5-year OS for drug-resistant recurrent ovarian cancer patients (44 ± 11.07 months, 95% CI: 231.24–53.66 months) was significantly lower than that of drug-sensitive recurrent ovarian cancer patients (58 ± 3.97 months; 95% CI: 50.05–65.59 months; p = 0.024) TP53 was the most frequently mutated gene in both drug-resistant (9/10, 90%) and drug-sensitive recurrent ovarian cancers (10/11, 91%). MYC and RB1 had the highest frequency of copy number variations (6/21, 29%) in recurrent ovarian cancers, followed by PIK3CA (3/21, 14%). BRCA2 N372H polymorphism was found in 40% (4/10) of drug-resistant recurrent ovarian cancer patients. The specificity, sensitivity, and consistency of TP53 and BRCA1 in circulating tumor-free DNA and tumor tissue DNA were 100%, 73.7%, 76.2% and 100%, 75%, 95.24%, respectively. Conclusion: We uncovered extensive genetic alterations in recurrent ovarian cancer and drug-resistant recurrent ovarian cancer exhibited unique genetic changes compared with recurrent ovarian cancer and drug-sensitive recurrent ovarian cancer. We further showed that high-throughput sequencing using liquid biopsy specimens could provide an effective, specific, and sensitive approach for detecting genetic alterations in ovarian cancer.