The relationship between TACC3 overexpression in tumor cells and poor prognosis has been reported in several malignant tumors such as esophageal squamous cell carcinoma, nonsmall cell lung cancer, hepatocellular carcinoma, gastric cancer, and soft tissue sarcoma [11, 15, 19, 21, 36, 37]. However, little is known about the association of TACC3 overexpression in osteosarcoma with patients’ clinicopathologic features and outcome. Therefore, we assessed the potential of TACC3 as a prognostic predictor for osteosarcoma and found an association of TACC3 overexpression with metastasis-free survival and overall survival in a small group of patients with osteosarcoma.
There are some limitations of our results. First, the number of patients in this study was relatively small. In addition, not all patients with osteosarcoma had sufficient followup duration or complete enough records to analyze; there may have been transfer bias. The problem with having a small cohort is weakness of the ability to perform a valid multivariate analysis, which needs a large cohort. It was unusual that tumor necrosis after chemotherapy was not associated with prognosis. Second, immunohistochemical (IHC) analysis was the sole method of assessing TACC3 expression. Because TACC3 is involved in microtubule regulation, the expression of TACC3 should be expected in all cells undergoing mitosis. So, the IHC antibody may also mark abnormally expressed TACC3. Hence, we performed polymerase chain reaction-based TACC3 copy number assay to measure the association of TACC3 expression with gene amplification. However, we could not find the correlation between TACC3 protein expression and gene amplification (data not shown). More detailed analysis of a mutation or IHC target such as an inactive form of TACC3 would be necessary to solve the question of how TACC3 is abnormally expressed in patients with osteosarcoma.
TACC3 expression was observed in many mitotic tumor cells by IHC analysis. This is consistent with previous observations that TACC3 is expressed, especially in the mitotic phase [5, 6]. However, IHC analysis also detected TACC3 in nonmitotic tumor cells. In normal human tissues, TACC3 is expressed in highly proliferative tissues such as the testes, spleen, thymus, and peripheral blood lymphocytes [23, 28]. Additionally, it was reported that TACC3 expression is associated with the proliferative activity of hepatocellular carcinoma cells . In our study, higher TACC3 expression was correlated with that of Ki-67. This marker is expressed in the nuclei throughout the cell cycle, except for the G0 phase, and reflects the cell’s proliferative ability . These findings suggest that TACC3 is expressed in tumor cells with proliferative potential as well as in mitotic tumor cells. TACC3 expression was correlated with increased p53 expression, but with the number of patients we had, we could not show an association with high p53 expression and poor prognosis in this study. Opinions are divided regarding the relationship between high p53 expression and poor prognosis in patients with osteosarcoma, whereas many reports have demonstrated that high p53 expression is an indicator of poor prognosis in this disease .
Patients with high TACC3 expression had shorter metastasis-free survival and overall survival compared with patients with low TACC3 expression. Furthermore, after controlling for likely confounding variables, we found that high TACC3 expression was associated with an increased likelihood of metastasis and death in patients with osteosarcoma. There are several possible explanations for this. First, TACC3 upregulation could activate the PI3K/AKT and ERK signaling pathways, which promotes epithelial-mesenchymal transition. Epithelial-mesenchymal transition provides tumor cells with migratory ability and increases their invasive capacity . In our study, higher TACC3 expression was correlated with larger tumor size (ie, AJCC stage IIB in this study). The IIA and IIB subdivision in the AJCC stage is useful for predicting a subsequent metastatic event . Second, higher levels of TACC3 could decrease DNA repair ability, resulting in genomic instability . However, it is possible that p53-mediated apoptosis might not occur because the p53-dependent checkpoint of tumor cells may be compromised in the TACC3 (high)/p53 (high) group as in nonsmall cell lung cancer .
Several studies have demonstrated the relationship between high TACC3 expression in tumor cells and poor prognosis in various cancers [11, 15, 19, 21, 36, 37]. However, there is no consensus regarding the cutoff value for high TACC3 expression based on IHC analysis. Therefore, we determined the cutoff value for TACC3 expression by analyzing the ROC curve and Youden index using metastasis, an established indicator of osteosarcoma prognosis , as a dichotomous variable as well as TACC3 positivity rates as the continuous variable. In addition, the cutoff value for p53 and Ki-67 expression was also defined based on the ROC curve and Youden index. Our cutoff value using the ROC curve, and not an arbitrary value, is considered appropriate .
It has been reported that TACC3 disruption suppresses tumor growth without damaging normal cells , induces premature senescence in tumor cells , and increases the susceptibility of tumor cells to paclitaxel, which is a microtubule inhibitor . In addition, spindlactone (SPL) and Specific and Non-genetic IAP-dependent Protein Eraser (SNIPER) (TACC3) have been developed as TACC3 inhibitors [22, 34]. SPL suppresses the extension of the microtubule from the centrosome and causes mitotic defects in tumor cells by inhibiting TACC3, whereas SNIPER (TACC3) selectively induces apoptosis in tumor cells expressing increased levels of TACC3 by polyubiquitylation and proteasomal degradation. These studies indicate that TACC3 may represent a novel, promising therapeutic target, although the potential must be tested either in animal models or in proper clinical trials. In this study, we provide no evidence that these approaches will be successful.
In conclusion, we demonstrated for the first time that high TACC3 expression is correlated with poor prognosis in patients with osteosarcoma. Further studies about mutation or an inactive form of TACC3 may reveal how TACC3 is abnormally expressed in osteosarcoma. If substantiated in a larger study, the observations from this pilot study might provide the rationale to use TACC3 as a potential therapeutic target.
We thank Mayumi Miura, Kanoko Miyazaki, Yuki Morotomi, Chie Kuroki, Kaoruko Nagatomo, Marina Sakata, and Kensaku Sato for their technical support.
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