C4.4A is a glycolipid-anchored membrane protein expressed in several human malignancies. The aim of this study was to explore the association between C4.4A expression at the invasion front of colorectal cancer (CRC) and tumor budding, a putative hallmark of cell invasion of CRC. Advanced CRCs (T2-4, n = 126) had a budding count of 3.66 ± 5.66, which was significantly higher than that of T1 early CRCs (1.75 ± 2.78, n = 87). C4.4A-positive CRC specimens showed a larger budding cell number than C4.4A-negative CRC specimens in T1 CRCs, and especially advanced CRCs (9.45 ± 5.83 vs 1.60 ± 3.93). Furthermore, we found a correlation between the percentage of C4.4A-positive cases and budding count in advanced CRC. Multivariate analysis for patients' survival showed that C4.4A was superior to tumor budding as a prognostic factor. With siRNA treatment, C4.4A levels were associated with cell invasion, but not with proliferation, in HCT116 and DLD1 cell lines. An immunohistochemical study in a subset of CRCs showed no relationship between C4.4A and Ki-67 proliferation marker. In vitro assays using HCT116 indicated that C4.4A levels correlated well with epithelial-mesenchymal transition (EMT) with regard to cell morphology and alterations of EMT markers including E-cadherin, vimentin, and partially N-cadherin. We also found that C4.4A expression was significantly associated with loss of E-cadherin and gain of b-catenin in clinical CRC tissue samples. These findings suggest that a tight association between C4.4A and tumor budding may, in part, be due to C4.4A promoting EMT at the invasive front of CRC. (Cancer Sci 2012; 103: 1155-1164 T he C4.4A protein has been identified in a highly metastasizing rat pancreatic adenocarcinoma cell line but not in locally growing rat tumors.(1,2) Rat C4.4A cDNA was cloned in 1998, and the molecular structure indicates GPI-anchored membrane proteins with 30% homology to the urokinase-type plasminogen activator receptor.(3) Subsequently, the human homologue of rat C4.4A, located on chromosome 19q13.1-q13.2, was cloned in 2001.(4) In normal human tissues, C4.4A mRNA is present in placental tissue, skin, esophagus, and peripheral blood leukocytes but not in other tissues, based on Northern blot analysis.(4) Although the physiological function of the C4.4A protein is largely unknown, upregulation of C4.4A expression has been observed during the wound-healing process of migrating keratinocytes or urothelium. (5,6) Recent studies have shown that C4.4A expression is also present in subsets of human malignancies. Human C4.4A mRNA has been detected in cancer cell lines, including melanoma, breast, bladder, and renal cell carcinoma, as well as in tumor tissue samples from malignant melanoma, colorectal cancer (CRC), breast cancer, lung carcinoma, and urothelial tumors.(6-10) C4.4A expression increases in metastatic lymph nodes and metastasized skin lesions compared with primary malignant melanoma,and evidence suggests that C4.4A expression is associated with poor prognosis of patients with non-small-cel...
C4.4A is a glycolipid-anchored membrane protein expressed in several human malignancies. We recently found that C4.4A expression was associated with poor prognosis of esophageal squamous carcinoma cells (ESCCs), but the underlying mechanism is unknown. To uncover this, we performed PCR array analysis using the HCT116 cell line, a positive control for C4.4A expression and we found that Tenascin-C (TNC) among the many adhesion molecules and extracellular matrix proteins was the best candidate for C4.4A molecule induction. Based on in vitro studies using the TE8 esophageal cancer cells, we examined by immunohistochemistry TNC expression in 111 ESCCs. We found that the TNC-positive group (24.3%) had significantly poorer prognosis than the TNC-negative group in 5-year overall survival. We also found there was a significant correlation between TNC and C4.4A in ESCC tissues (P=0.007). Finally, we found that only the double-positive group for C4.4A and TNC had a significantly worse prognosis (P=0.005). Our data suggest that TNC expression in ESCC may in part explain why C4.4A is associated with a poor prognosis of ESCC since TNC can promote invasion and metastasis.
Abstract. Intra-abdominal desmoid tumor is a lifethreatening disease. Studies have shown that dacarbazine (DTIC)-doxorubicin (DOX) (D-D) therapy is the most effective treatment. However, myelosuppression is a major problem, and cardiac muscle disorders due to DOX limit the number of administration cycles, whereas it usually requires a long time to achieve tumor shrinkage. To resolve these issues, we introduced low-dose D-D therapy to 3 patients employing 50 mg/m 2 DOX and 600-700 mg/m 2 DTIC per cycle, which permits repeated administration cycles up to 10-11 times. Case 1 was a 23-year-old female with a sporadic recurrent mesenterium desmoid tumor located in the pelvis (maximum diameter, 8 cm). Cases 2 and 3 were a 33-year-old female and a 36-year-old male. Both patients had intra-abdominal mesenterium desmoid tumors (maximum diameter 9.6 and 9.0 cm, respectively) that were generated after proctocolectomy due to familial adenomatous polyposis. No severe adverse events occurred during the therapy. With the aid of sulindac and tamoxifen after low-dose D-D therapy, the first two patients achieved a complete response, and the third patient achieved a partial response and awaits further tumor shrinkage. Our experience indicates that low-dose DT-D therapy is a safe and effective regimen for patients with intra-abdominal desmoid tumors.
The metastasis-associated gene C4.4A encodes a glycolipid-anchored membrane protein expressed in several human malignancies. The present study aimed to perform a detailed assessment of C4.4A expression in colorectal cancer tissues, in terms of intra-cellular localization, intra-tumoral location and difference in molecular weight. To advance this goal, we developed three new antibodies against the C4.4A protein (two polyclonal Abs: C4.4A-119 and C4.4A-277 and one monoclonal Ab: C4.4A GPI-M) to use in addition to the two previously produced polyclonal Abs (C4.4A-81, C4.4A GPI-P). Antibody specificities were confirmed by absorption tests. Western blot analysis and immunohistochemistry showed that the C4.4A-119 and C4.4-277 Abs detected 70-kDa C4.4A, mainly in the cytoplasm, irrespective of intra-tumoral location. The C4.4A GPI-P and C4.4A GPI-M Abs reacted with the membranous ~40-kDa C4.4A, exclusively at the tumor invasive front, and each detected an identical tumor cell population. The tested antibodies showed varied C4.4A detection rates in 33 CRC tissues. The C4.4A-277 Ab yielded the highest positive rate in 29 of 33 CRC tissues (87.9%), while the C4.4A GPI-P and C4.4A GPI-M Abs each only showed 33.3% positivity. The present findings suggest that the GPI anchor signaling sequence may be essential for detecting membranous C4.4A at the invasive front of CRC tissues.
Following the publication of this article, an interested reader drew to our attention an anomaly associated with the presentation of the Fig. 2D; specifically, there appeared to have been a duplication of the same β-actin control band for the G4.4A GPI-m and the Absorbed Ab (antibody) experiments.After having re-examined our original data, we realize that the same β-actin control bands were inadvertently selected for the data shown in Fig. 2D, and this escaped out attention at the time. Our investigation of the data has also confirmed that the β-actin bands in the lower panels were produced from simultaneously performed blots using different gels or lanes with the same HCT116 cell lysates.A corrected version of Fig. 2, containing alternative data obtained from an experiment performed in duplicate, is showin below. The error with the selection of the control data did not affect the results in this study. We sincerely apologize for this mistake, and thank the reader of our article who drew this matter to our attention. Furthermore, we regret any inconvenience this mistake has caused. Figure 2. Western blot analysis for the C4.4A protein using lysates from HCT116 colon cancer cells. (A) C4.4A-119 antibody; (B) C4.4A-277 antibody; (C) C4.4A GPI-P antibody; and (D) C4.4A GPI-m antibody. The arrows indicate the band corresponding to the C4.4A protein. β-actin bands in the lower panels were produced from simultaneously performed western blot analyses using different gels or lanes with the same lysates from HCT116 cells.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.
Copyright © 2025 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.