Hong Liu

The application of CRISPR/Cas9 system in cervical carcinogenesis

AbstractIntegration of high-risk HPV genomes into cellular chromatin has been confirmed to promote cervical carcinogenesis, with HPV16 being the most prevalent high-risk type. Herein, we evaluated the therapeutic effect of the CRISPR/Cas9 system in cervical carcinogenesis, especially for cervical precancerous lesions. In cervical cancer/pre-cancer cell lines, we transfected the HPV16 E7 targeted CRISPR/Cas9, TALEN, ZFN plasmids, respectively. Compared to previous established ZFN and TALEN systems, CRISPR/Cas9 has shown comparable efficiency and specificity in inhibiting cell growth and colony formation and inducing apoptosis in cervical cancer/pre-cancer cell lines, which seemed to be more pronounced in the S12 cell line derived from the low-grade cervical lesion. Furthermore, in xenograft formation assays, CRISPR/Cas9 inhibited tumor formation of the S12 cell line in vivo and affected the corresponding protein expression. In the K14-HPV16 transgenic mice model of HPV-driven spontaneous cervical carcinogenesis, cervical application of CRISPR/Cas9 treatment caused mutations of the E7 gene and restored the expression of RB, E2F1, and CDK2, thereby reversing the cervical carcinogenesis phenotype. In this study, we have demonstrated that CRISPR/Cas9 targeting HPV16 E7 could effectively revert the HPV-related cervical carcinogenesis in vitro, as well as in K14-HPV16 transgenic mice, which has shown great potential in clinical treatment for cervical precancerous lesions.

Clinical efficacy analysis of chemotherapy of isolated neck lymphatic metastasis in advanced epithelial ovarian cancer

The aim of this study was to retrospectively investigate the efficacy of chemotherapy for neck lymph node metastasis (NLNM) by determining the characteristics and survival of patients with isolated NLNMs metastases from epithelial ovarian carcinoma (EOC) at stage IV of the Federation of Gynecology and Obstetrics (FIGO). The clinicopathological characteristics and survival outcome of 24 cases with stage IV FIGO EOC with isolated NLNM were retrospectively analyzed between December 1, 2014, and November 30, 2021. Among the 24 patients, 2 (8.3%) underwent primary debulking surgery (PDS), 21 (87.5%) received neoadjuvant chemotherapy(NACT) followed by interval debulking surgery (IDS), and 1 (4.2%) received chemotherapy alone. Additionally, 13 (54.2%) cases achieved abdominal R0 debulking, while 11(45.8%) cases achieved R1/R2 debulking. The chemotherapy response of NLNMs included complete response (8/24, 33.3%), partial response (15/24,62.5%), or stable disease (1/24,41.7%). None of the patients received resection or radiotherapy of NLNMs. Recurrence was observed in 15 (62.5%) patients, with only 2 experiencing recurrence of NLNMs. The median progression-free survival (PFS) and overall survival (OS) were 35 months and 48 months, respectively. R0 debulking led to a significantly longer PFS (not reached) and OS (57 months) compared to non-R0 debulking (PFS: 10 months, P = 0.001; OS: 22 months, P = 0.001). Interestingly, patients with EOC with lymphatic recurrence had better OS ( 57 months) than did those with abdominal or distant recurrence (OS: 29 months; P = 0.012). Chemotherapy is an effective treatment for neck lymph nodes metastasis, and a favorable response to chemotherapy could eliminate the necessity for NLNM resection or radiotherapy. Effective control of abdominal disease with surgery may be a critical factor in managing FIGO stage IV EOC patients with isolated NLMNs.