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@#[摘 要] 目的：探讨lncRNA ZNF674-AS1在宫颈癌中的作用及其分子机制。方法：用qPCR法检测ZNF674-AS1在31例2019年1月至2020年7月在武汉儿童医院接受手术治疗患者的宫颈癌组织和对应的癌旁组织、宫颈癌细胞系（SiHa、HeLa、C33A和HCC94）和永生化子宫颈上皮细胞系中的表达。转染过表达ZNF674-AS1质粒及其阴性对照质粒至ZNF674-AS1表达最少的HCC94细胞，CCK-8法和Transwell实验检测过表达ZNF674-AS1对HCC94细胞增殖活性和迁移能力的影响。生物信息学方法预测和双荧光素酶报告基因实验验证ZNF674-AS1和miR-510-5p、REPS2三者间的互补结合关系。qPCR检测过表达ZNF674-AS1对miR-510-5p与REPS2表达的影响，WB法检测过表达ZNF674-AS对细胞增殖和迁移相关因子表达的影响。结果：与癌旁组织相比，ZNF674-AS1在宫颈癌组织中呈明显低表达（P<0.01）；与永生化子宫上皮细胞相比，ZNF674-AS1在宫颈癌细胞系中也呈明显低表达（P<0.05或P<0.01），以HCC94细胞中表达最低（P<0.01）。过表达ZNF674-AS1能明显抑制HCC94细胞的增殖（P<0.05）和迁移（P<0.01）。与ZNF674-AS1相互作用的miRNA是miR-510-5p，与miR-510-5p相互作用的基因是REPS2。过表达ZNF674-AS1导致HCC94细胞中miR-510-5p的表达水平降低（P<0.01）而REPS2基因的表达水平升高（P<0.01），同时引起细胞增殖和迁移相关的多种因子（CDK2、cyclin D3、vimentin和twist）上调或下调。结论：lncRNA ZNF674-AS1在宫颈癌组织和细胞中呈低表达，可能通过竞争性结合miR-510-5p而上调REPS2的表达，从而抑制宫颈癌HCC94细胞的增殖和迁移。
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@#[Abstract] Objective: To investigate the effect of HMGB1 gene on the growth of human epithelial ovarian cancer xenografts in nude mice, and to lay a foundation for finding new targets for the treatment of ovarian cancer. Methods: Human epithelial ovarian cancer SKOV3 cells in logarithmic growth phase were selected to establish a human epithelial ovarian cancer xenograft model in nude mice. Nude mice with successful model establishment were randomly divided into control group and HMGB1-siRNA group. On the 7th, 9th, 11th, 14th, and 16th days after cell inoculation, the same amount of saline and HMGB1-siRNA were respectively injected into two groups of mice under the armpit.After 3 weeks, the nude mice were sacrificed by cervical dislocation, the tumor tissues were separated, and the volume of the tumor was measured. The apoptosis of transplanted tumor cells was detected by Tunnel staining. The expressions of HMGB1, STAT3 and p-STAT3 were detected by Western blotting. The expression of vascular endothelial growth factorA(VEGF-A) and microvascularization were detected by immunohistochemistry. Results: Compared with the control group, the growth of tumor volume slowed down in HMGB1 siRNA group, and on the 21st day, the tumor volume of HMGB1-siRNA group was significantly smaller than that of the control group (P<0.05). HMGB1-siRNA successfully knocked down the expression of HMGB1 mRNA in transplanted tumor tissue. The apoptosis rate of tissue cells in HMGB1-siRNA group was significantly increased ([34±8]% vs [6±2]%, P=0.04), and the expressions of HMGB1 and p-STAT3 were significantly reduced (P<0.05). The expression of VEGF-Aand the number of microvessels were significantly lower than those of the control group (both P<0.05). Conclusion: Knockdown of HMGB1 gene reduces the expression of VEGF-A and microvessel formation possibly by inhibiting the HMGB1/STAT3 signaling pathway, thereby promoting the apoptosis of tumor tissues and slowing the growth of xenografts.
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@# Objective: To investigate whether miR-140 could increase the sensitivity of cervical cancer (CC) to oxaliplatin by downregulating the expression of programmed death-1 (PD-L1). Methods: qPCR was used to analyze miR-140 expression in normal human cervical cells, CC cells and oxaliplatin-resistant CC cells. Cells were transfected with miR-140 mimic, and then, the proliferation of CC cells and oxaliplatin-resistant CC cells was detected by using CCK-8 assay, and the colony formation rate of CC cells was obtained by using colony formation assay. Starbase and TargetScan were used to predict the targeted binding site of miR-140 and PD-L1, and the influence of miR-140 on the expression of PD-L1 was validated by dual luciferase reporter gene assay.Annexin V FITC/PI double staining and Wb assays were used to detect the effect of over-expression of miR-140 or both over-expression of PD-L1 and miR140 on the apoptosis, migration and expression of apoptosis-related proteins in CC cells after treatment with oxaliplatin. Moreover, transplantation tumor of CC cell lines was established in nude mice to assess the effects of miR-140 on enhancing the sensitivity of tumors to oxaliplatin. Results: The expression of miR-140 was significantly decreased in oxaliplatin-resistant CC cells (P<0.01). Over-expression of miR140 could significantly increase the sensitivity of oxaliplatin-resistant CC cells to oxaliplatin (P<0.05), and inhibit the CC cells proliferation and colony formation (P<0.01). miR-140 showed targeted binding to PD-L1 3'-UTR and inhibited its expression. Over-expression of miR-140 significantly promoted CC cell migration and apoptosis (P<0.01). However, co-transfection of PD-L1 counteracts the effects of miR-140 on cell metastasis and apoptosis (all P<0.05). In addition, xenograft tumor model in mice also verified that miR-140 could promote the sensitivity of tumors to oxaliplatin. Conclusion: miR-140 increases the sensitivity of CC to oxaliplatin through inhibition of PD-L1 expression. Therefore, up-regulation of miR-140 or down-regulation of PD-L1 in combination with oxaliplatin may be a novel strategy for the treatment of Oxaliplatin-resistant CC.