睾丸特异蛋白Y编码样5 (TSPYL5) 在体内抑制多种癌症，包括结直肠癌（CRC）;然而，其在体内对CRC细胞肿瘤发生的机制和作用仍不明确。为阐明结直肠癌的分子机制并寻找新的治疗靶点以改善CRC患者的预后，我们使用四周龄、16-22克的雄性小鼠放置在温度控制房间（20-25°C）的无菌笼中，12小时的光照/黑暗循环，以及自由摄食并获得饮水。使用TSPYL5 过表达或非过表达的HCT116 细胞构建裸鼠肿瘤模型。用苏木精与伊红(H&E)染色对肿瘤组织进行组织学评估。TUNEL染色评估肿瘤细胞凋亡。免疫组化法测量切除的肿瘤组织中的Ki67表达。通过免疫印迹法检测体内双链断裂（DBS）相关蛋白的表达。使用IBM SPSS统计软件（Windows版, 版本21.0) 对所有分析进行。至少进行三个独立实验得出均值±标准差。使用非配对学生t检验比较组别。使用单因素方差分析（one-way ANOVA）和Dunnett's检验比较P<0.5的组。TSPYL5过表达抑制CRC细胞的肿瘤形成能力并且损伤肿瘤细胞在体内。TSPYL5过表达还显著增加Bax和p-H2AX（早期双链断裂指示物）的表达，并降低Ki67、Bcl-2和过氧化物酶起动子-激活受体的表达。总之，TSPYL5过表达通过诱导DNA损伤来抑制CRC细胞的肿瘤形成能力。

Testis-specific protein Y-encoded-like 5 (TSPYL5) suppresses several cancers in vivo, including colorectal cancer (CRC); however, its mechanism and role in CRC cell tumorigenesis in vivo remain unknown.To elucidate the molecular mechanisms of colorectal cancer and find new therapeutic targets to improve CRC patient outcomes.Male mice (4 weeks old, 16-22 g) were housed in sterile cages in a temperature-controlled room (20-25°C) with a 12 h light/dark cycle and ad libitum food and water.TSPYL5 overexpressing or non-overexpressing HCT116 cells were used to create a nude mouse tumor model. Tumor tissue was evaluated histologically after hematoxylin and eosin (H and E) staining. TUNEL staining assessed tumor cell apoptosis. Ki67 expression in excised tumor tissue was measured by immunohistochemistry. Western blotting examined double-stranded break (DBS)-associated protein expression in vivo.IBM SPSS Statistics for Windows, Version 21.0 was used for all analyses (IBM Corp., Armonk, NY, USA). At least three independent experiments yield a mean value ± standard deviation. Unpaired Student's t-tests compared groups. One-way analysis of variance and Dunnett's test were used to compare groups with a P value < 0.5.TSPYL5 overexpression inhibited CRC cell tumorigenicity and damaged tumor cells in vivo. TSPYL5 overexpression also significantly increased Bax and p-H2AX (early double-stranded break indicators) and decreased Ki67, Bcl-2, and peroxisome proliferator-activated receptor expression.Collectively, TSPYL5 overexpression inhibited the tumorigenicity of CRC cells in vivo by inducing DNA damage.