Yajie Wang

Genetic silencing of CDC6 via AAV2-Delivered shRNA as a novel cancer genetics–based therapy for cervical carcinoma

Cell division cycle 6 (Cdc6) is an oncogenic driver in cervical cancer, whose dysregulation accelerates S-phase entry and promotes genomic instability. As a key replication licensing factor, its overexpression creates a cancer-specific vulnerability, making it a promising therapeutic target. To evaluate whether silencing Cdc6 via an adeno-associated virus serotype 2 (AAV2)-delivered shRNA can selectively inhibit cervical cancer growth while sparing normal cells. We constructed an AAV2 vector encoding short hairpin RNA (shRNA) targeting Cdc6 and validated its efficacy in vitro using multiple cervical cancer cell lines and an immortalized epithelial cell line (HaCaT). Functional assays assessed cell cycle progression, apoptosis, and DNA damage. Antitumor efficacy was further assessed in xenograft mouse models. AAV2-shCdc6 transduction efficiently silenced Cdc6 expression, leading to G2/M phase arrest, increased γ-H2AX expression, and significant apoptosis in cervical cancer cells. In contrast, normal HaCaT cells exhibited only S-phase arrest without apoptosis. In vivo, AAV2-shCdc6 treatment significantly inhibited tumor growth in xenograft models without observable systemic toxicity. AAV2-mediated Cdc6 knockdown selectively targets cervical cancer by exploiting a defined genetic vulnerability. This cancer genetics-based strategy offers a precise and well-tolerated approach for cervical cancer therapy.

KIF18A Is a Novel Target of JNK1/c‐Jun Signaling Pathway Involved in Cervical Tumorigenesis

ABSTRACTCervical cancer remains a significant global health concern. KIF18A, a kinesin motor protein regulating microtubule dynamics during mitosis, is frequently overexpressed in various cancers, but its regulatory mechanisms are poorly understood. This study investigates KIF18A's role in cervical cancer and its regulation by the JNK1/c‐Jun signaling pathway. Cell growth was assessed in vitro using MTT and colony formation assays, and in vivo using a nude mouse xenograft model with KIF18A knockdown HeLa cells. The Genomic Data Commons (GDC) data portal was used to identify KIF18A‐related protein kinases in cervical cancer. Western blot analysis was employed to analyze phosphor‐c‐Jun, c‐Jun, and KIF18A expression levels following JNK1 inhibition, c‐Jun knockdown/overexpression, and KIF18A knockdown in cervical cancer cells. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were performed to assess c‐Jun binding and transcriptional activity of the KIF18A promoter. KIF18A knockdown significantly impaired cervical cancer cell growth both in vitro and in vivo. A strong positive correlation was observed between JNK1 and KIF18A expression in cervical and other cancers. JNK1 inhibition decreased both KIF18A expression and c‐Jun phosphorylation. c‐Jun was found to directly bind to and activate the KIF18A promoter. Furthermore, c‐Jun knockdown inhibited cervical cancer cell growth, and this effect was partially rescued by KIF18A overexpression. This study demonstrates that the JNK1/c‐Jun pathway activates KIF18A expression, which is essential for cervical cancer cell growth. Targeting the JNK/c‐Jun/KIF18A axis may represent a promising novel therapeutic strategy for cancer treatment.