Activation of the hedgehog pathway in human hepatocellular carcinomas

Liver cancers, the majority of which are hepatocellular carcinomas (HCCs), rank as the fourth in cancer mortality worldwide and are the most rapidly increasing type of cancer in the United States. However, the molecular mechanisms underlying HCC development are not well understood. Activation of the hedgehog pathway is shown to be involved in several types of gastrointestinal cancers. Here, we provide evidence to indicate that hedgehog signaling activation occurs frequently in HCC. We detect expression of Shh, PTCH1 and Gli1 in 115 cases of HCC and in 44 liver tissues adjacent to the tumor. Expression of Shh is detectable in about 60% of HCCs examined. Consistent with this, hedgehog target genes PTCH1 and Gli1 are expressed in over 50% of the tumors, suggesting that the hedgehog pathway is frequently activated in HCCs. Of five cell lines screened, we found Hep3B, Huh7 and PLC/PRF/5 cells with detectable hedgehog target genes. Specific inhibition of hedgehog signaling in these three cell lines by smoothened (SMO) antagonist, KAAD-cyclopamine, or with Shh neutralizing antibodies decreases expression of hedgehog target genes, inhibits cell growth and results in apoptosis. In contrast, no effects are observed after these treatments in HCC36 and HepG2 cells, which do not have detectable hedgehog signaling. Thus, our data indicate that hedgehog signaling activation is an important event for development of human HCCs.     

The sonic hedgehog signaling pathway is reactivated in human renal cell carcinoma and plays orchestral role in tumor growth

Background  

Human clear cell renal cell carcinoma (CRCC) remains resistant to therapies. Recent advances in Hypoxia Inducible Factors (HIF) molecular network led to targeted therapies, but unfortunately with only limited clinical significance. Elucidating the molecular processes involved in kidney tumorigenesis and resistance is central to the development of improved therapies, not only for kidney cancer but for many, if not all, cancer types. The oncogenic PI3K/Akt, NF-kB and MAPK pathways are critical for tumorigenesis. The sonic hedgehog (SHH) signaling pathway is crucial to normal development.     

Suppressed MKP-1 is an independent predictor of outcome in patients with hepatocellular carcinoma

OBJECTIVE: An increase in the activity of mitogen-activated protein kinases (MAPKs) has been correlated with a more malignant phenotype in several tumor models in vivo. This study was designed to clarify the expression of MKP-1 in surgically resected hepatocellular carcinoma (HCC). METHODS: We reviewed the cases of 77 patients who had undergone initial liver resection for HCC without preoperative treatment. Immunohistochemical analysis of MKP-1 was performed on paraffin-embedded tissues. The correlation between MKP-1 expression and clinical outcome was investigated. RESULTS: Tumor cells were immunohistochemically stained for MKP-1 expression, and the same levels as in normal hepatocytes were detected in 66 (85%) of 77 HCC patients, being decreased in 11 (15%) HCCs. Decreased MKP-1 expression significantly correlated with serum alpha-fetoprotein levels and tumor size (p<0.05). The disease-free survival rates in MKP-1-negative and -positive patients were 0 and 31.0% at 5 years, respectively (p<0.01). The survival rates after a surgical resection in MKP-1-negative and -positive patients were 18.2 and 65.5% at 5 years, respectively (p<0.01). CONCLUSIONS: The MKP-1 expression in HCC was an independent prognostic factor for outcome in HCC patients. In the future, it will be useful to explore whether the phosphatase expression might account for the response to HCC treatments targeting at MAPK activation.     

Serum insulin-like growth factor-1 is an independent predictor of prognosis in patients with renal cell carcinoma

Obesity is associated with an increased risk of certain cancers, including renal cell carcinoma. A possible mediator of this risk is insulin-like growth factor-1 (IGF-1). The authors evaluated the prognostic information of IGF-1, IGFBP-3, leptin, and prealbumin in sera sampled at diagnosis from 256 consecutive patients with renal cell carcinoma. Insulin-like growth factor-1 and leptin were positively correlated to body mass index (BMI). Insulin-like growth factor-1 and IGFBP-3 did not correlate to tumour stage or grade. Leptin and prealbumin were both inversely related to tumour stage and grade. When survival was analysed in patients with levels above a median of IGF-1, leptin, and prealbumin, prognosis was more favourable, compared with those with lower levels (p=0.017; p=0.024, and p<0.0001, respectively). In a multivariate analysis, tumour stage and serum IGF-1 levels were independent prognostic factors. The results indicate that serum IGF-1 at diagnosis is related to prognosis in renal cell carcinoma.     

The mTOR pathway is associated with the poor prognosis of human hepatocellular carcinoma

Objective The mammalian target of rapamycin (mTOR) pathway, an important regulator of multiple cellular functions including proliferation, differentiation, tumorigenesis, and apoptosis, is up-regulated in many cancers. It has achieved considerable importance. This study was conducted to determine the status of the mTOR pathway in human hepatocellular carcinoma (HCC) and to investigate its relationship with the prognosis of HCC. Methods PTEN, pAkt, p27, and pS6 expression in cryo-sections gathered from 528 cases with HCC by the method of immunohistochemistry. Kaplan–Meier survival and Cox regression analyses were performed to evaluate the prognosis of HCC. Results The mTOR pathway was more significantly altered in high-grade tumors, and tumors with poor prognostic features. Especially, pAkt and cytoplasmic p27 expression showed the strongest associations with pathological parameters of HCC. Statistical analysis showed that HCC patients expressing pAkt, PTEN, cytoplasmic p27, and pS6 have different overall survival rates relative to those not expressing these proteins. Cox multi-factor analysis showed that tumor differentiation (P = 0.006), vascular invasion (P = 0.028), TNM stage (P = 0.005), pAkt (P = 0.021), PTEN (P = 0.003), p27 (P = 0.018) and pS6 (P = 0.002) were independent prognosis factors for HCC. Conclusion: Expression of the mTOR pathway components, which are related with the transferability and invasive capacity of HCC cells, may be used as prognostic indicators in HCC.     

Persistent elevated C-reactive protein after treatment is an independent marker of a poor prognosis in patients with hepatocellular carcinoma

Purpose

The pretreatment C-reactive protein (CRP) level is reported to be a prognostic indicator in patients with hepatocellular carcinoma (HCC).

Methods

We investigated the prognostic implications of the changes in the CRP level after initial treatment in patients with HCC. We prospectively evaluated a cohort of 150 patients with newly diagnosed HCC. The patients were categorized into three groups: group 1 (n = 120) with pre- and post-treatment CRP <1.0 mg/dl, group 2 (n = 5) with pre-treatment CRP ≥1.0 mg/dl and post-treatment CRP <1.0 mg/dl, and group 3 (n = 25) with pre- and post-treatment CRP ≥1.0 mg/dl.

Results

The 1- and 3-year overall survival rates were 92.3 and 82.9 % for group 1, 80.0 and 53.3 % for group 2, and 58.8 and 4.2 % for group 3. The overall survival rate for group 3 was significantly lower than that for group 1 (P < 0.0001), or group 2 (P = 0.003). No significant difference was found between groups 1 and 2 (P = 0.627). A multi-variate analysis showed that albumin level (P = 0.049), the CRP group (P < 0.0001), and the Cancer of the Liver Italian Program (CLIP) score (P < 0.0001) were independently associated with the overall survival.

Conclusions

A persistently elevated CRP level after initial treatment is an independent marker of a poor prognosis, and normalization of the CRP level after initial treatment is associated with a better outcome in patients with HCC.     

TGFBI expression is an independent predictor of survival in adjuvant-treated lung squamous cell carcinoma patients

Background:

Transforming growth factor β-induced protein (TGFBI) is a secreted protein that mediates cell anchoring to the extracellular matrix. This protein is downregulated in lung cancer, and when overexpressed, contributes to apoptotic cell death. Using a small series of stage IV non-small cell lung cancer (NSCLC) patients, we previously suggested the usefulness of TGFBI as a prognostic and predictive factor in chemotherapy-treated late-stage NSCLC. In order to validate and extend these results, we broaden the analysis and studied TGFBI expression in a large series of samples obtained from stage I–IV NSCLC patients.

Methods:

TGFBI expression was assessed by immunohistochemistry in 364 completely resected primary NSCLC samples: 242 adenocarcinomas (ADCs) and 122 squamous cell carcinomas (SCCs). Kaplan–Meier curves, log-rank tests and the Cox proportional hazards model were used to analyse the association between TGFBI expression and survival.

Results:

High TGFBI levels were associated with longer overall survival (OS, P<0.001) and progression-free survival (PFS, P<0.001) in SCC patients who received adjuvant platinium-based chemotherapy. Moreover, multivariate analysis demonstrated that high TGFBI expression is an independent predictor of better survival in patients (OS: P=0.030 and PFS: P=0.026).

Conclusions:

TGFBI may be useful for the identification of a subset of NSCLC who may benefit from adjuvant therapy.     

Long-term outcome of patients with insular carcinoma of the thyroid: the insular histotype is an independent predictor of poor prognosis

BACKGROUND: Insular thyroid carcinoma was described originally as a tumor with aggressive behavior. However, whether a predominant insular component is an independent factor for poor prognosis is unclear. METHODS: The authors compared the clinical behavior of tumors in three groups of patients with thyroid carcinoma--13 patients with insular thyroid carcinoma, 18 patients with follicular thyroid carcinoma, and 26 patients with papillary thyroid carcinoma--who were selected based on similar tumor size and similar age. Disease free survival and disease specific deaths were assessed in the three groups with a Kaplan-Meier analysis and were compared using the log-rank test. Cox regression analysis was used to evaluate the influence of histotype and other prognostic factors on the occurrence of distant metastases and disease specific death. RESULTS: Patient follow-up ranged from 5.2 months to 190.0 months. At last follow-up, only 1 of 13 patients (7.7%) with insular carcinoma, compared with 8 of 18 patients (44.4%) with follicular carcinoma and 12 of 26 patients (46.1%) with papillary carcinoma, were disease free. The disease specific death rate was 61.5% among patients in the insular carcinoma group compared with 16.7% and 15.4% among patients in the follicular carcinoma group (P = 0.006) and the papillary carcinoma group (P = 0.025), respectively. At multivariate analysis, the insular histotype was the only variable that was related independently to disease specific death (hazard ratio = 4.27; P = 0.005). Distant metastases occurred in 84.6% of patients in the insular carcinoma group compared with 50% and 19.2% of patients in the follicular carcinoma group (P = 0.039) and the papillary carcinoma group (P = 0.0003), respectively. All metastases from patients with insular carcinomas (n = 11 patients) showed radioiodine uptake, but a clinical benefit from this treatment was observed only in 1 patient. CONCLUSIONS: Patients with insular thyroid carcinoma have a poorer outcome compared with patients of similar age who have differentiated types of thyroid carcinoma with tumors of a similar size. Because radioiodine rarely is effective in the treatment of patients with metastatic insular thyroid carcinoma, novel and possible multimodal therapies should be explored for the treatment of patients with these aggressive tumors.     

High preoperative plasma fibrinogen is an independent predictor of distant metastasis and poor prognosis in renal cell carcinoma

Background

We aimed to evaluate the association of preoperative plasma fibrinogen levels with the clinicopathological parameters, disease-free survival, and overall survival in patients with renal cell carcinoma.

Methods

We retrospectively studied 286 patients with renal cell carcinoma who underwent radical nephrectomy from 2000 to 2003 at one center. The plasma fibrinogen was routinely determined before operation in all patients. The correlation of preoperative plasma fibrinogen levels with clinicopathological findings was evaluated by t-test or analysis of variance (ANOVA) methods. As well, univariate and multivariate analyses were used to determine the association between the preoperative level of plasma fibrinogen and survival duration.

Results

An elevated level of plasma fibrinogen was positively related to the Fuhrman grade (P < 0.001), tumor size (P < 0.001), and T stage (P < 0.001), but it was negatively related to histologic type (P = 0.266). Univariate analysis showed that the Fuhrman grade, tumor size, T stage, hemoglobin, corrected calcium, lactate dehydrogenase, and plasma fibrinogen level were significantly correlated with disease-free survival (P < 0.001, P < 0.001, P < 0.001, P < 0.001, P = 0.001, P < 0.001, and P < 0.001, respectively) and overall survival (P < 0.001, P = 0.001, P < 0.001, P < 0.001, P = 0.002, P = 0.001, and P < 0.001). Multivariate analysis showed that the plasma fibrinogen level remained as an independent prognostic factor for disease-free survival (P = 0.021) and overall survival (P < 0.001).

Conclusions

A high preoperative plasma fibrinogen level is an independent predictor of distant metastasis and survival prognosis after radical nephrectomy in patients with renal cell carcinoma.     

Desmoglein‐2 expression is an independent predictor of poor prognosis patients with multiple myeloma

Multiple myeloma (MM) is the second most common haematological malignancy and is an incurable disease of neoplastic plasma cells (PC). Newly diagnosed MM patients currently undergo lengthy genetic testing to match chromosomal mutations with the most potent drug/s to decelerate disease progression. With only 17% of MM patients surviving 10‐years postdiagnosis, faster detection and earlier intervention would unequivocally improve outcomes. Here, we show that the cell surface protein desmoglein‐2 (DSG2) is overexpressed in ~ 20% of bone marrow biopsies from newly diagnosed MM patients. Importantly, DSG2 expression was strongly predictive of poor clinical outcome, with patients expressing DSG2 above the 70^th percentile exhibiting an almost 3‐fold increased risk of death. As a prognostic factor, DSG2 is independent of genetic subtype as well as the routinely measured biomarkers of MM activity (e.g. paraprotein). Functional studies revealed a nonredundant role for DSG2 in adhesion of MM PC to endothelial cells. Together, our studies suggest DSG2 to be a potential cell surface biomarker that can be readily detected by flow cytometry to rapidly predict disease trajectory at the time of diagnosis.     

Activation of the hedgehog pathway in chronic myelogeneous leukemia patients

Background

Here we aimed to investigate the effect of COX-2 siRNA on proliferation and angiogenesis of gastric cancer cells.

Methods

The gastric cancer cell line SGC7901 was transfected with COX-2 siRNA, then the growth and angiogenesis of cells were detected by in vitro and in vivo assay. Human microarray, RT-PCR and western blot were used to identify differentially expressed angiogenesis-related molecules in cells with decreased expression of COX-2.

Results

Down-regulation of COX-2 could significantly inhibit the in vitro and in vivo growth of gastric cancer cells, and suppress the migration and tube formation of human umbilical vein endothelial cells. Totally 23 angiogenesis-related molecules were found involved in COX-2-induced angiogenesis suppression. The results of RT-PCR and western blot showed that down-regulation of COX-2 might inhibit VEGF, Flt-1, Flk-1/KDR, angiopoietin-1, tie-2, MMP2 and OPN.

Conclusions

COX-2 might mediate tumor angiogenesis and growth, and could be considered as a target for gastric cancer therapy.     

Tissue inhibitor of metalloproteinase 1 is an independent predictor of prognosis in patients with nonsmall cell lung carcinoma who undergo resection with curative intent

BACKGROUND: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play a role in the processes of extracellular matrix degradation. Changes in their expression levels have been observed in various tumor types, including lung carcinoma. However, their clinical significance and their prognostic importance in the progression of nonsmall cell lung carcinoma (NSCLC) remain to be specified. In this study, mRNA expression levels of MMP-1, MMP-9, TIMP-1, and TIMP-2 were evaluated in patients with resected NSCLC, and their associations with disease progression and prognosis were determined. METHODS: Between June 1996 and December 1999, 116 patients underwent resection for NSCLC. Expression levels of MMPs and TIMPs were evaluated using Northern blot analysis in these NSCLC tissue samples and in 39 matched samples of normal lung tissue. RESULTS: MMP-1, MMP-9, and TIMP-1 expression levels were increased in tumor samples compared with matched, corresponding normal tissues. In contrast, TIMP-2 expression was decreased in tumor samples. MMP-1 tumor expression was correlated significantly with the evolution of lymph node status and tumor-lymph node-metastasis (TNM) stage. In contrast, MMP-9 tumor expression was correlated significantly with increased T stage. TIMP-1 overexpression was an independent predictor of worse survival in patients with NSCLC that was not associated with other prognosis factors, such as TNM stage. CONCLUSIONS: The overexpression of TIMP-1 was an independent prognostic marker in patients with NSCLC, and evaluating TIMP-1 may be important for identifying patients who are at greater risk of disease recurrence.     

Activation of hedgehog signaling is not a frequent event in ovarian cancers

The hedgehog (Hh) signaling pathway regulates many processes of development and tissue homeostasis. Activation of hedgehog signaling has been reported in about 30% of human cancer including ovarian cancer. Inhibition of hedgehog signaling has been pursued as an effective strategy for cancer treatment including an ongoing phase II clinical trial in ovarian cancer. However, the rate of hedgehog signaling activation in ovarian cancer was reported differently by different groups. To predict the successful for future clinical trials of hedgehog signaling inhibitors in ovarian cancer, we assessed hedgehog pathway activation in 34 ovarian epithelial tumor specimens through analyses of target gene expression by in-situ hybridization, immunohistochemistry, RT-PCR and real-time PCR. In contrast to previous reports, we only detected a small proportion of ovarian cancers with hedgehog target gene expression, suggesting that identification of the tumors with activated hedgehog signaling activation will facilitate chemotherapy with hedgehog signaling inhibitors.     

Tamoxifen is not effective in good prognosis patients with hepatocellular carcinoma

Background  

Large randomised clinical trials and systematic reviews substantiate that tamoxifen is ineffective in improving survival of patients with hepatocellular carcinoma (HCC). However, a recent report suggested that the drug might prolong survival among patients with well preserved liver function. The aim of this paper is to validate this hypothesis.