利用体内CRISPR/Cas9全基因组敲除文库筛选发现DUSP4抑制宫颈癌淋巴结转移的作用与机制研究

Lymph nodes metastasis , which plays significant role in the prognosis and treatment decision in cervical cancer patients, is also the most common metastatic way in this disease. Therefore, unveiling the mechanism behind this process and exploring new target for treatment can be of great importance. we performed in vivo genome-wide CRISPR/Cas9 knockout screening to identify important regulators of cervical cancer lymph node metastasis. we found that the DUSP family member,DUSP4 and DUSP5, stood out as the top hits in the screening.We confirmed these findings in cervical cancer patient samples. Notably, expression of DUSP4 was dramatically decreased in lymph node positive cervical cancer samples and the deficiency of DUSP4 enhances metastasis of cervical cancer cell in vitro,These data suggest that DUSP4 might play an important role in regulating cervical cancer lymph node metastasis. We next investigated the molecular mechanism of DUSP4 deficiency in promoting cervical cancer metastasis by using RNA-seq, Immunoprecipitation and LC-MS .The notable finding is that interaction between DUSP4 and YAP1 might be a novel mechanism in control cervical cancer lymph node metastasis, we validated the interaction between DUSP4 and YAP1 by Co-IP assays, and we want to take more in-depth studies to uncover the underlying mechanisms and to find out whether YAP1 inhibitor can serve as a potential therapeutic strategy for the treatment of metastatic cervical cancer. Our findings will improve our understanding of DUSP4 and YAP1 roles and their cross-talk in regulation of cervical cancer lymph node metastasis, which further contributes to improve the current strategy for cervical cancer therapy.

宫颈癌淋巴结转移是最常见的转移途径以及影响患者预后的重要因素，揭示其机制并探寻新的治疗靶点是其有效治疗的关键。我们利用体内CRISPR/Cas9全基因组敲除文库来发现调控宫颈癌淋巴结转移的关键基因，我们发现了两个双特异性磷酸酶家族基因：DUSP4和DUSP5最为显著，并在宫颈癌临床样本和细胞实验中进行了验证。我们发现DUSP4在淋巴结阳性患者中的表达显著下调，且DUSP4的失活会促进宫颈癌细胞的转移。这些数据提示我们DUSP4可能是抑制淋巴结转移的关键基因。接下来我们通过RNA测序，免疫沉淀联合质谱分析来进一步探索。发现DUSP4与YAP1的相互作用可能是一种新的调控淋巴结转移的分子机制。我们希望通过更多实验来验证表型，发现具体分子调控机制以及YAP1抑制剂能否成为新的治疗选择。本课题开展将加深我们对DUSP4与YAP1互作调控淋巴转移的了解，为宫颈癌淋巴结转移的治疗提供新的策略。

宫颈癌淋巴结转移是最常见的转移途径以及影响患者预后的重要因素，揭示其机制并探寻 新的治疗靶点是其有效治疗的关键。我们利用体内CRISPR/Cas9全基因组敲除文库来发现调控 宫颈癌淋巴结转移的关键基因，我们发现了两个双特异性磷酸酶家族基因：DUSP4和DUSP5最为显著，并在宫颈癌临床样本和细胞实验中进行了验证。我们发现DUSP4在淋巴结阳性患者中的表达显著下调，且DUSP4的失活会促进宫颈癌细胞的转移。这些数据提示我们DUSP4可能是抑制淋巴结转移的关键基因。通过免疫沉淀联合质谱分析，我们发现DUSP4和AXL之间存在相互作用。我们通过进一步实验验证了DUSP4-AXL信号通路调控宫颈癌淋巴结转移的分子机制，并发现AXL的抑制剂能够明显抑制宫颈癌淋巴结转移。本研究不仅为宫颈癌淋巴结转移机制提供理论基础，也为今后宫颈癌转移治疗提供新方向。