Transcription factor BATF3 promotes malignant biological behavior of clear cell renal cell carcinoma cells by regulating vimentin / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨碱性亮氨酸拉链转录因子ATF样蛋白3（BATF3）在肾透明细胞癌（ccRCC）组织中的表达及其调控ccRCC细胞恶性生物学行为的分子机制。方法：收集2019年3月至2022年1月间在中国人民解放军联勤保障部队第九八〇医院手术切除的64例ccRCC组织和相应癌旁组织，qPCR法检测ccRCC组织、癌旁组织和肾癌ACHN、786-O细胞中BATF3 mRNA的表达，免疫组化检测ccRCC组织、癌旁组织中BATF3蛋白的表达，并分析其与临床病理特征的关系。构建BATF3敲减及过表达质粒，分别转染786-O、ACHN细胞，通过MTS法、Transwell实验检测BATF3对786-O或ACHN细胞增殖、迁移、侵袭能力的影响，qPCR法检测敲减或过表达BATF3对786-O或ACHN细胞EMT相关基因表达的影响，CHIP、双荧光素酶报告基因实验检测BATF3是否与波形蛋白（VIM）启动子区结合并调控其转录，MTS法、Transwell实验检测同时过表达BATF3及敲减VIM对786-O细胞增殖、迁移、侵袭能力的影响。结果：与癌旁组织比较，ccRCC组织中BATF3的mRNA和蛋白均呈高表达(均P<0.01)，并且BATF3 mRNA与ccRCC的分化程度和TNM分期密切关联（均P<0.01）；与正常肾上皮细胞293T相比，BATF3在ACHN及786-O细胞中均呈高表达（均P<0.01）。敲减BATF3表达均能明显抑制786-O细胞的增殖、迁移、侵袭能力(均P<0.01)，过表达BATF3则均能促进ACHN细胞的增殖、迁移和侵袭能力（均P<0.01），敲减或过表达BATF3能抑制786-O细胞或促进ACHN细胞的EMT相关基因的表达（均P<0.01）。BATF3可与VIM启动子区的位点结合，直接调控VIM的转录表达。同时过表达BATF3及敲减VIM可逆转过表达BATF3对786-O细胞增殖、迁移、侵袭能力的影响。结论：BATF3在ccRCC组织中呈高表达，并与其分化程度和TNM分期密切关联；BATF3通过调控VIM的表达影响ACHN、786-O细胞的恶性生物学行为，其可作为临床治疗ccRCC的潜在靶点。

Expression of forkhead box protein D3 in gastric cardia adenocarcinoma tissues and its effect on biological behaviors of SGC-7901 cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨叉头框蛋白D3（forkhead box protein D3，FOXD3）在胃贲门腺癌（gastric cardia adenocarcinoma，GCA）中的表达及其对SGC-7901细胞生物学行为的影响。方法：从河北医科大学第四医院生物标本库中选取2014年6月至2016年12月手术切除的49例GCA组织及相应癌旁组织标本，qRT-PCR检测FOXD3在GCA组织、癌旁组织以及在5种胃癌细胞系中（BGC-823、SGC-7901、HGC-27、MGC-803及NCI-N87）的表达。向SGC-7901细胞转染pc-DNA3.1-FOXD3或pc-DNA3.1，采用细胞增殖实验、克隆形成实验、划痕愈合实验和Transwell小室侵袭实验分别检测FOXD3过表达对SGC-7901细胞增殖、克隆形成、迁移和侵袭的影响，qRT-PCR及WB法检测细胞转染前后上皮-间质转化（epithelial-mesenchymal transition，EMT）相关分子mRNA及蛋白的表达情况，流式细胞术检测转染前后细胞周期改变。结果：GCA组织中FOXD3 mRNA的表达量明显降低，其表达水平与患者临床分期和淋巴结转移密切关联；FOXD3在胃癌细胞系中的表达均低于正常细胞（均P<0.01）。FOXD3过表达能明显抑制SGC-7901细胞的增殖、克隆形成、迁移和侵袭能力（均P<0.01），提高SGC-7901细胞中E-cadherin的表达水平，减少N-cadherin、β-catenin和vimentin的表达水平（均P<0.01），使细胞周期阻滞在G0/G1期（P<0.01）。结论: FOXD3在GCA组织中的表达明显下调，其过表达可以抑制胃癌细胞的生物学行为，FOXD3可作为抑癌基因为肿瘤治疗提供新思路。

TGF- β induced lncRNA linc01503 promotes proliferation, invasion and EMT process of esophageal squamous cell carcinoma cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of lncRNA linc01503 in esophageal squamous cell carcinoma (ESCC) tissues and cell lines, as well as its effect on the proliferation, invasion and epithelial-mesenchymal transition (EMT) of ESCC cells. Methods: A total of 119 pairs of tumor tissues and corresponding para-cancerous tissues of ESCC patients were obtained from the Fourth Affiliated Hospital of Hebei Medical University between Jan. 2012 and Dec. 2014. The expression of linc01503 in ESCC tissues and cell lines (Kyse150, Kyse170, Eca109, TE1 and TE13) was detected by qPCR. The ESCC cells were transfected with pGPU6-shRNA-linc01503 or treated with TGF-β. The expressions of EMT related genes before and after transfection as well as linc01503 expression before and after TGF-β treatment were detected with qPCR. MTS and Transwell assay were performed to assess the effect of linc01503 on proliferation and invasion of ESCC cells. Results: The expression of linc01503 was significantly elevated in ESCC tissues and cell lines (all P<0.05). High expression of linc01503 was correlated with lymph node metastasis, depth of infiltration, TNM stage and the survival of ESCC patients (all P<0.05). Treatment with TGF-β promoted EMT of ESCC cells and induced a significant up-regulation of linc01503 expression. Knockdown of linc01503 significantly inhibited proliferation and invasion ability of ESCC cells; Meanwhile, the low expression of linc01503 increased the expression of E-cadherin but decreased the expressions of N-cadherin and vimentin (all P<0.05). Conclusion: lncRNA linc01503, as one of the downstream effect genes of TGF- β, promotes the proliferation, invasion and EMT process of ESCC cells.

Effects of PTPN6 on malignant biological behaviors of human esophageal squamous cancer Eca109 andYes-2 cells / 中国肿瘤生物治疗杂志

@#Objective: To detect the expression of non-receptor protein tyrosine phosphatase 6 (PTPN6) in different esophageal squamous cell carcinoma (ESCC) cell lines, and to investigate its effect on proliferation, migration and invasion ability of Eca109 and Yes-2 cell lines. Methods: qPCR was applied to detect the mRNA expression of PTPN6 in different ESCC cell lines (TE1, Eca109, Kyse150, Kyse170 and Yes-2). pcDNA3.1-PTPN6 plasmid was transiently transfected into Eca109 and Yes-2 cells respectively. The expression of PTPN6 was detected by real-time PCR and Wb. The effects of PTPN6 over-expression on the biological behaviors of ESCC cells were detected by MTS, colony formation assay, wound healing assay and Transwell assay, respectively. Results: The mRNA expression of PTPN6 was remarkably reduced in ESCC cell lines (TE1, Eca109, Kyse150, Kyse170 and Yes-2) compared to normal esophageal epithelial cells (HEEpiC) (P<0.05). Compared to the mock cells, significant up-regulation of PTPN6 was detected in pcDNA3.1-PTPN6 transfected Eca109 and Yes-2 cells (all P<0.05 or P<0.01); PTPN6 over-expression led to a significant inhibition in migration and invasion ability of Eca109 and Yes-2 cells (all P<0.05). Conclusions: Over-expression of PTPN6 may inhibit the proliferation, migration and invasion of ESCC cells, which might be an important factor influencing the biological characteristics of ESCC cells.