背景：在全球范围内，结直肠癌（Colorectal cancer, CRC）的发病率及死亡率均位于恶性肿瘤前列，准确的预后判断是提高患者生存率的关键。目前，TNM 分期被广泛用于 CRC 的肿瘤分期及预后预测。然而，CRC 在分子水平上存在高度异质性，临床病理特征相同的患者预后可能存在显著差异，因此生物标志物有望在提高 CRC 患者预后判断准确性方面发挥重要作用。本研究旨在评估 DNAJB6、KIAA1522 和 p-mTOR 蛋白的表达水平在 CRC 预后中的价值，并建立有效的 CRC 患者预后模型。

方法：首先采用免疫组化方法检测 329 例 CRC 标本中 DNAJB6、KIAA1522 和 p-mTOR（Ser2448）蛋白的表达，分析其表达量与临床病理参数的关系。接下来将患者随机分为训练队列（n = 230）及验证队列（n = 99），采用 Kaplan-Meier 法、单因素和多因素 Cox 比例风险回归模型评估训练队列中的三种蛋白的预后价值，并构建了整合三种蛋白和 TNM 分期的预后列线图模型。随后，我们使用受试者工作特征（Receiver operating characteristic， ROC）曲线、一致性指数（The concordance index， C-index）、校准曲线和决策曲线分析（Decision curve analysis，DCA）评估列线图在训练和验证队列中的预测效能。

结果：DNAJB6、KIAA1522 和 p-mTOR 三个蛋白在 CRC 组织中的表达显著上调（P < 0.01），其表达水平是总体生存期（Overall survival，OS）和无病生存期 （Disease-free survival，DFS）的独立预后因素（P 均 < 0.05）。ROC 曲线下面积和一致性指数约为 0.7。校准曲线结果表明模型预测值与实际观测值的一致性较好。DCA 曲线表明列线图模型的临床效益高于 TNM 分期。总体而言，该预测模型具有满意的一致性、敏感性与特异性及临床实用性。

结论：DNAJB6、KIAA1522 和 p-mTOR 高表达是 CRC 预后不良的独立影响因子。整合  DNAJB6、KIAA1522 和 p-mTOR 及 TNM 分期的列线图可以准确提供 CRC 患者的预后信息，有望成为 CRC 预后的新型监测工具。

背景：结直肠癌（colorectal cancer，CRC）是世界范围内流行的消化系统恶性肿瘤，发病率及死亡率均居前列。肿瘤细胞向周围组织侵袭转移是晚期患者预后不良的重要原因，发现调控结直肠癌侵袭转移的关键基因并对其进行生物学功能的分析，已成为目前 CRC 领域的研究热点。

目的：探讨微管相关蛋白4（MAP4）对 CRC 细胞恶性表型的影响及相关分子机制。

方法：利用免疫组织化学方法检测 CRC 组织中 MAP4 蛋白表达量的变化，采用卡方检验及 Fisher 精确检验探索 MAP4 蛋白表达与 CRC 患者临床病理参数的相关性。采用 Kaplan-Meier 法及 Cox 风险比例回归模型探索 MAP4 表达水平与 CRC 患者预后的相关性。在结肠癌细胞系 DLD1 和 HCT116 中转染小干扰 RNA（siRNA）和短发夹 RNA（shRNA）敲降 MAP4 表达 ；在稳定敲降 MAP4 的 HCT116 及 DLD1 中瞬时转染 MAP4 同义突变的过表达质粒载体 （标记为 MAP4-R）进行回复实验。利用 CCK 8 细胞增殖活力实验、细胞集落形成实验、伤口愈合实验、Transwell 实验检测 MAP4 对 DLD1 和 HCT116 细胞恶性表型的影响；利用 Western blot 检测 MAP4 及相关下游分子的蛋白表达改变。

结果：使用含有 496 例 CRC 患者组织的微阵列进行免疫组织化学检测，结果显示 MAP4 在 CRC 组织中显著上调（P < 0.001）。结合相关临床病理资料进行统计分析发现， MAP4 表达水平与淋巴结转移 （P = 0.009）、局部浸润深度（P < 0.001）、组织病理学分级（P < 0.001）以及病理 TNM 分期（P = 0.002）显著正相关；使用Cox 比例风险模型进行多因素回归分析，结果提示 MAP4 是 CRC 患者的独立预后因素（总生存期，P = 0.015；无病生存期，P = 0.020）。进而，我们选取 MAP4 高表达的 CRC 细胞株 DLD1 和 HCT116 探究 MAP4 异常表达对 CRC 细胞恶性表型的影响。体外功能研究结果显示，敲降 MAP4 表达后 CRC 细胞的增殖能力无明显改变，但伤口愈合能力和侵袭迁移能力显著降低。回复实验提示在稳定敲降 MAP4 的细胞中外源过表达 MAP4 后，细胞的侵袭迁移能力得到恢复。分子水平检测发现干扰 MAP4 表达可导致 ERK 和 S6 的磷酸化水平明显下调。

结论：MAP4 在 CRC 中异常高表达并且是患者 OS 及 DFS 的独立预测因素，同时其高表达可能通过激活 ERK 及 S6 相关信号通路增强 CRC 细胞的侵袭迁移能力。

Background: Globally, the incidence and mortality of colorectal cancer (CRC) are at the forefront of malignant tumors, and accurate prognosis judgment is the key to improve the survival rate of patients. At present, TNM stage system is widely used for tumor staging and prognosis prediction of CRC. However, CRC is highly heterogeneous at the molecular level, and there may be significant differences in the prognosis of patients with the same clinicopathological features, so molecular markers are expected to play an important role in improving the accuracy of prognosis in patients with CRC. This study aims to evaluate the value of expression levels of DNAJB6, KIAA1522, and p-mTOR proteins in the prognosis of CRC and to establish a valid prognostic model for patients with CRC.

Methods: Firstly, the expression of DNAJB6, KIAA1522, and p-mTOR (Ser2448) proteins in 329 CRC specimens was detected by immunohistochemistry, and the relationship between their expression and clinicopathological parameters was analyzed. Next, the patients were randomly divided into a training set (n = 230) and validation set (n = 99), and the prognostic value of the three proteins in the training set was evaluated by the Kaplan-Meier method, univariate and multivariate Cox proportional hazard regression models, and the prognostic nomograms model integrating the three proteins and TNM stage system were constructed. Subsequently, we used the Receiver operating characteristic (ROC) curve, The concordance index (C-index), the calibration curve, and the Decision curve analysis (DCA) to evaluate the predictive power of the nomogram in the training and validation cohorts.

Results: The expression of DNAJB6, KIAA1522, and p-mTOR in CRC tissues was significantly upregulated (P < 0.01), and their expression levels were independent prognostic factors for overall survival (OS) and disease-free survival (DFS) (both P < 0.05). The area under the ROC curve and the C-index values are approximately 0.7. The findings of the calibration curve demonstrate that the model's projected values and actual measurements coincide rather well. The DCA curve indicates that the nomogram model has a higher clinical benefit than the TNM stage system. Overall, the predictive model has satisfactory consistency, sensitivity, specificity, and clinical practicability.

Conclusion: High expression of DNAJB6, KIAA1522, and p-mTOR are independent influencing factors for the poor prognosis of colorectal cancer. Nomograms integrating DNAJB6, KIAA1522, p-mTOR, and TNM stage systems can accurately provide prognostic information for colorectal cancer patients and are expected to become new monitoring tools for colorectal cancer prognosis.

Background: Colorectal cancer (CRC) is a world-wide epidemic malignant tumor of the digestive system, with the highest incidence and mortality. The invasion and metastasis of tumor cells to surrounding tissues is an important reason for the poor prognosis of advanced patients, and the discovery of key genes that regulate colorectal cancer invasion and metastasis and the analysis of their biological functions have become a research hotspot in the field of CRC.

Objective: To investigate the effect of microtubule-associated protein 4 (MAP4) on the malignant phenotype of colorectal cancer cells and related molecular mechanisms.

Methods: Immunohistochemistry was used to detect the change of MAP4 protein expression in CRC tissues, and Chi-square test and Fisher precision test were used to explore the correlation between MAP4 protein expression and clinicopathological parameters in CRC patients. The Kaplan-Meier method and Cox risk proportional regression model were used to explore the correlation between MAP4 expression level and the prognosis of CRC patients. Transfect small interfering RNA (siRNA) and short hairpin RNA (shRNA) to colorectal cancer cell  DLD1 and HCT116 by silencing MAP4. Recovery experiments were performed in HCT116 and DLD1 that stably knocked down MAP4 with an overexpressed plasmid vector (labeled MAP4-R) synonymously. The effects of  MAP4 on the malignant phenotype of DLD1 and HCT116 cells were detected by CCK8 cell proliferation and viability experiments, cell colony formation experiments, wound healing experiments, and Transwell experiments. Western blot was used to detect changes in the expression of MAP4 and related downstream molecular proteins.

Results: Tissue microarray and immunohistochemical testing in 496 colorectal cancer patients showed significant upregulation of MAP4 in colorectal cancer tissues (P < 0.001). We found that MAP4 expression was associated with local lymph node metastasis (P = 0.009), local invasion depth (P < 0.001), and histopathological grade (P < 0.001) and pathologic TNM stage (P = 0.002) were closely related, and Cox proportional hazards regression analysis suggested that MAP4 may be an independent prognostic factor in colorectal cancer patients (overall survival, P = 0.015; disease-free survival, P = 0.020). Next, we selected DLD1 and HCT116 colorectal cancer cell lines with high expression of MAP4 to explore the effect of MAP4 on the malignant phenotype of colorectal cancer, and found that the proliferation ability of cells did not change significantly after silencing MAP4, the wound healing and invasion migration ability of tumor cells were significantly reduced. The response experiment showed that after exogenous overexpression of MAP4 in cells with stable knockdown of MAP4, the invasion and migration ability of cells was restored. Molecular level testing has found that interference with MAP4 can lead to downregulation of phosphorylation levels of ERK and S6.

Conclusion: MAP4 is abnormally high expressed in CRC and is an independent predictor of OS and DFS in patients, and its high expression may enhance the invasion and migration capacity of CRC cells by activating ERK and S6-related signaling pathways.