pFAK-Y397 overexpression as both a prognostic and a predictive biomarker for patients with metastatic osteosarcoma.

Focal adhesion kinase (FAK) is important for tumor cell survival and metastasis in various cancers. However, its expression and prognostic value in patients with metastatic osteosarcoma remain unknown. We investigated the expression of FAK and its phosphorylated form (pFAK-Y397) in osteosarcoma tissues from 53 patients by immunohistochemistry and evaluated their correlations with clinicopathologic characteristics and outcomes. The prognostic values were assessed using Kaplan-Meier survival and Cox regression analyses. Total FAK and pFAK-Y397 were overexpressed in 48 (90.6%) and 33 (62.3%) cases, respectively. pFAK-Y397 overexpression was correlated with poor histologic response after neoadjuvant chemotherapy in patients with osteosarcoma regardless of the presence of metastasis or not. Kaplan-Meier curve showed that patients with metastatic osteosarcoma with pFAK-Y397 overexpression had significantly worse overall survival (OS) than those with non-overexpression (P = 0.044). Multivariate Cox regression analysis confirmed pFAK-Y397 overexpression as an independent prognostic predictor for OS and post metastases OS (PMOS) (P = 0.017, P = 0.006, respectively). Age at diagnosis was also an independent indicator for PMOS (P = 0.003). However, total FAK expression was not correlated with any clinicopathologic characteristics or OS in patients with metastatic osteosarcoma. In conclusion, our findings identified FAK as a common aberrant protein overexpression in various subtypes of osteosarcoma. pFAK-Y397 overexpression can be used as a prognostic biomarker predicting poor OS for patients with metastatic osteosarcoma, and the expression of pFAK-Y397 differentiated good and poor responders to neoadjuvant chemotherapy.

PTEN Expression as a Predictor of Response to Focal Adhesion Kinase Inhibition in Uterine Cancer.

PTEN is known to be frequently mutated in uterine cancer and also dephosphorylates FAK. Here, we examined the impact of PTEN alterations on the response to treatment with a FAK inhibitor (GSK2256098). In vitro and in vivo therapeutic experiments were carried out using PTEN-mutated and PTEN-wild-type models of uterine cancer alone and in combination with chemotherapy. Treatment with GSK2256098 resulted in greater inhibition of pFAK(Y397) in PTEN-mutated (Ishikawa) than in PTEN-wild-type (Hec1A) cells. Ishikawa cells were more sensitive to GSK2256098 than the treated Hec1A cells. Ishikawa cells were transfected with a wild-type PTEN construct and pFAK(Y397) expression was unchanged after treatment with GSK2256098. Decreased cell viability and enhanced sensitivity to chemotherapy (paclitaxel and topotecan) in combination with GSK2256098 was observed in Ishikawa cells as compared with Hec1a cells. In the Ishikawa orthoptopic murine model, treatment with GSK2256098 resulted in lower tumor weights and fewer metastases than mice inoculated with Hec1A cells. Tumors treated with GSK2256098 had lower microvessel density (CD31), less cellular proliferation (Ki67), and higher apoptosis (TUNEL) rates in the Ishikawa model when compared with the Hec1a model. From a large cohort of evaluable patients, increased FAK and pFAK(Y397) expression levels were significantly related to poor overall survival. Moreover, PTEN levels were inversely related to pFAK(Y397) expression. These preclinical data demonstrate that PTEN-mutated uterine cancer responds better to FAK inhibition than does PTEN wild-type cancer. Therefore, PTEN could be a biomarker for predicting response to FAK-targeted therapy during clinical development.

Age and survival of cervical cancer patients with bone metastasis.

BACKGROUND: To determine survival times of cervical cancer patients with bone metastasis related to the effect of age at the time of cervical cancer diagnosis, we performed the retrospectively analytical study. METHODS: A total of 68 cervical cancer patients with bone metastasis were treated at a single hospital, during January 1998 to December 2010. Fifty-two medical records were identified and collected, the remaining sixteen medical records were not found. Main outcome measures were patient characteristics, clinical information, duration from cervical cancer diagnosis to bone metastasis diagnosis, survival time after bone metastasis and overall survival time. RESULTS: Among fifty-two cervical cancer patients with bone metastasis, there were 13 patients who were less than 45 years old, and 39 patients were 45 years old or more at the time of cervical cancer diagnosis. The younger group had less median overall survival than the older group, with a statistically significant difference (21 months, 95% CI 19.93-22.06; 34 months, 95% CI 23.27-44.72, p = 0.021). However, they were comparable in the duration from cervical cancer diagnosis to bone metastasis diagnosis and the survival time after bone metastasis. CONCLUSION: Young patients with bone metastasis aged less than 45 years old at the time of cervical cancer diagnosis have a poorer prognosis than the elderly patients. IMPACT: To improve survival and quality of life, more intensive and novel multimodal treatments at the time of cervical cancer diagnosis should be considered in patients less than forty-five years, who can tolerate the side effects better.

Focal adhesion kinase: an alternative focus for anti-angiogenesis therapy in ovarian cancer.

This investigation describes the clinical significance of phosphorylated focal adhesion kinase (FAK) at the major activating tyrosine site (Y397) in epithelial ovarian cancer (EOC) cells and tumor-associated endothelial cells. FAK gene amplification as a mechanism for FAK overexpression and the effects of FAK tyrosine kinase inhibitor VS-6062 on tumor growth, metastasis, and angiogenesis were examined. FAK and phospho-FAK(Y397) were quantified in tumor (FAK-T; pFAK-T) and tumor-associated endothelial (FAK-endo; pFAK-endo) cell compartments of EOCs using immunostaining and qRT-PCR. Associations between expression levels and clinical variables were evaluated. Data from The Cancer Genome Atlas were used to correlate FAK gene copy number and expression levels in EOC specimens. The in vitro and in vivo effects of VS-6062 were assayed in preclinical models. FAK-T and pFAK-T overexpression was significantly associated with advanced stage disease and increased microvessel density (MVD). High MVD was observed in tumors with elevated endothelial cell FAK (59%) and pFAK (44%). Survival was adversely affected by FAK-T overexpression (3.03 vs 2.06 y, P = 0.004), pFAK-T (2.83 vs 1.78 y, P<0.001), and pFAK-endo (2.33 vs 2.17 y, P = 0.005). FAK gene copy number was increased in 34% of tumors and correlated with expression levels (P<0.001). VS-6062 significantly blocked EOC and endothelial cell migration as well as endothelial cell tube formation in vitro. VS-6062 reduced mean tumor weight by 56% (P = 0.005), tumor MVD by 40% (P = 0.0001), and extraovarian metastasis (P<0.01) in orthotopic EOC mouse models. FAK may be a unique therapeutic target in EOC given the dual anti-angiogenic and anti-metastatic potential of FAK inhibitors.

Biologic effects of platelet-derived growth factor receptor α blockade in uterine cancer.

PURPOSE: Platelet-derived growth factor receptor α (PDGFRα) expression is frequently observed in many kinds of cancer and is a candidate for therapeutic targeting. This preclinical study evaluated the biologic significance of PDGFRα and PDGFRα blockade (using a fully humanized monoclonal antibody, 3G3) in uterine cancer. EXPERIMENTAL DESIGN: Expression of PDGFRα was examined in uterine cancer clinical samples and cell lines, and biologic effects of PDGFRα inhibition were evaluated using in vitro (cell viability, apoptosis, and invasion) and in vivo (orthotopic) models of uterine cancer. RESULTS: PDGFRα was highly expressed and activated in uterine cancer samples and cell lines. Treatment with 3G3 resulted in substantial inhibition of PDGFRα phosphorylation and of downstream signaling molecules AKT and mitogen-activated protein kinase (MAPK). Cell viability and invasive potential of uterine cancer cells were also inhibited by 3G3 treatment. In orthotopic mouse models of uterine cancer, 3G3 monotherapy had significant antitumor effects in the PDGFRα-positive models (Hec-1A, Ishikawa, Spec-2) but not in the PDGFRα-negative model (OVCA432). Greater therapeutic effects were observed for 3G3 in combination with chemotherapy than for either drug alone in the PDGFRα-positive models. The antitumor effects of therapy were related to increased apoptosis and decreased proliferation and angiogenesis. CONCLUSIONS: These findings identify PDGFRα as an attractive target for therapeutic development in uterine cancer.

Cross-talk between EphA2 and BRaf/CRaf is a key determinant of response to Dasatinib.

PURPOSE: EphA2 is an attractive therapeutic target because of its diverse roles in cancer growth and progression. Dasatinib is a multikinase inhibitor that targets EphA2 and other kinases. However, reliable predictive markers and a better understanding of the mechanisms of response to this agent are needed. EXPERIMENTAL DESIGN: The effects of dasatinib on human uterine cancer cell lines were examined using a series of in vitro experiments, including MTT, Western blot analysis, and plasmid transfection. In vivo, an orthotopic mouse model of uterine cancer was utilized to identify the biologic effects of dasatinib. Molecular markers for response prediction and the mechanisms relevant to response to dasatinib were identified by using reverse phase protein array (RPPA), immunoprecipitation, and double immunofluorescence staining. RESULTS: We show that high levels of CAV-1, EphA2 phosphorylation at S897, and the status of PTEN are key determinants of dasatinib response in uterine carcinoma. A set of markers essential for dasatinib response was also identified and includes CRaf, pCRaf(S338), pMAPK(T202/Y204) (mitogen-activated protein kinase [MAPK] pathway), pS6(S240/244), p70S6k(T389) (mTOR pathway), and pAKT(S473). A novel mechanism for response was discovered whereby high expression level of CAV-1 at the plasma membrane disrupts the BRaf/CRaf heterodimer and thus inhibits the activation of MAPK pathway during dasatinib treatment. CONCLUSIONS: Our in vitro and in vivo results provide a new understanding of EphA2 targeting by dasatinib and identify key predictors of therapeutic response. These findings have implications for ongoing dasatinib-based clinical trials.

Molecular therapy as a future strategy in endometrial cancer.

Of all gynecologic cancers, endometrial cancer is the most common cancer in the US and Europe. In addition, it is presently the second most common gynecologic cancer in the world. As a result of increasing menopausal, obese and tamoxifen use women, the incidence of the cancer seems to be on the increase. Surgery is the major treatment, whereas postoperative radiation therapy in high-intermediate risk patients many prevent locoregional recurrence. Adjuvant chemotherapy can improve progression free survival in advanced or recurrent cancers. Molecular targeted therapies are now a focus of attention including anti-vascular endothelial growth factor (VEGF), mammalian target of rapamycin (mTOR) inhibitor and tyrosine kinase inhibitor (TKI). They may provide useful future strategies for control of endometrial malignancies in developing countries and across the world.

Targeted endometrial cancer therapy as a future prospect.

Among female-specific cancers worldwide, endometrial cancer is the third most common after breast cancer and cervical cancer. In addition, it is the most common gynecological cancer in the USA and Europe. The incidence of this disease appears to be increasing. The cause of this increase is multifactorial, but a few possible factors involved are increasing obesity, an aging population leading to more postmenopausal women and greater tamoxifen use. Surgery is generally the primary treatment of this disease and postoperative radiation therapy in some patients with high or intermediate risk may prevent locoregional recurrences. Adjuvant chemotherapy improves progression-free survival in advanced or recurrent cancer. However, overall survival in patients with advanced disease is poor. Hence, better therapy is needed and targeted molecular therapies are emerging as possible treatment candidates. These include molecules that target VEGF, mTOR, tyrosine kinases, human EGF receptors and FGF receptors. Therapies targeting specific molecular features should be evaluated in future strategies in the treatment of endometrial cancer.

Metronomic activity of CD44-targeted hyaluronic acid-paclitaxel in ovarian carcinoma.

PURPOSE: Most primary human ovarian tumors and peritoneal implants, as well as tumor vascular endothelial cells, express the CD44 family of cell surface proteoglycans, the natural ligand for which is hyaluronic acid. Metronomic dosing, the frequent administration of chemotherapeutics at substantially lower than maximum tolerated doses (MTD), has been shown to result in reduced normal tissue toxicity and to minimize "off-treatment" exposure resulting in an improved therapeutic ratio. EXPERIMENTAL DESIGN: We tested the hypothesis that hyaluronic acid (HA) conjugates of paclitaxel (TXL; HA-TXL) would exert strong antitumor effects with metronomic (MET) dosing and induce antiangiogenic effects superior to those achieved with MTD administration or with free TXL. Female nude mice bearing SKOV3ip1 or HeyA8 ovarian cancer cells were treated intraperitoneally (i.p.) with MET HA-TXL regimens (or MTD administration) to determine therapeutic and biologic effects. RESULTS: All MET HA-TXL-treated mice and the MTD group revealed significantly reduced tumor weights and nodules compared with controls (all P values < 0.05) in the chemotherapy-sensitive models. However, the MTD HA-TXL-treated mice showed significant weight loss compared with control mice, whereas body weights were not affected in the metronomic groups in HeyA8-MDR model, reflecting reduced toxicity. In the taxane-resistant HeyA8-MDR model, significant reduction in tumor weight and nodule counts was noted in the metronomic groups whereas the response of the MTD group did not achieve significance. While both MTD and metronomic regimens reduced proliferation (Ki-67) and increased apoptosis (TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling), only metronomic treatment resulted in significant reductions in angiogenesis (CD31, microvessel density). Moreover, metronomic treatment resulted in substantial increases in thrombospondin-1 (Tsp-1), an inhibitor of angiogenesis. CONCLUSIONS: This study showed that MET HA-TXL regimens have substantial antitumor activity in ovarian carcinoma, likely via a predominant antiangiogenic mechanism.

Moving beyond VEGF for anti-angiogenesis strategies in gynecologic cancer.

Gynecologic cancer is a major burden in both developed and developing countries. Almost a half million deaths from gynecologic cancer are reported each year. Understanding the molecular biology of cancer is a principle resource leading to the identification of new potential therapeutic targets, which may be parlayed into novel therapeutic options in gynecologic cancer. Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase, which plays a pivotal role in many aspects of malignant growth including cancer cell survival, migration, invasion, angiogenesis and metastasis. Various human cancer tissues have demonstrated high expression of FAK or activated FAK, which has been correlated with survival of cancer patients. Among gynecologic cancers, reports have emerged demonstrating that FAK is involved in the pathogenesis of ovarian, endometrial, and cervical cancers. In addition, the polycomb group protein enhancer of Zeste homologue 2 (EZH2), Dll4/notch and EphA2 has also emerged as important regulators of endothelial cell biology and angiogenesis. Herein, we review the role of these new targets in tumor angiogenesis and the rationale for further clinical development.

Knowledge of Pap smear, HPV and the HPV vaccine and the acceptability of the HPV vaccine by Thai women.

AIMS: The primary prevention for cervical cancer, a human papilloma virus (HPV) vaccine, has been available in Thailand for almost 3 years. The present study evaluates knowledge about the Papanicolaou (Pap) smear, HPV and the HPV vaccine and focuses on identifying predictors for the acceptability of the HPV vaccine. METHODS: A sample of 764 women attending the gynecology clinic at Ramathibodi Hospital, Bangkok, was asked to answer a questionnaire on their personal background, their knowledge of the Pap smear and HPV and the HPV vaccine and the acceptability of the HPV vaccine for themselves and their daughters. RESULTS: Knowledge of the Pap smear (96%) is higher than that of HPV (41%) and the HPV vaccine (36%). Only 40% of participants had previously heard about HPV. The acceptability of the HPV vaccine for participants and their daughters was high, 77% and 84%, respectively. Knowing about HPV increases acceptance for the HPV vaccine (adjusted OR = 1.7, 95% CI = 1.2-2.5, in the participants and OR = 2.3, 95% CI = 1.5-3.6 in their daughters). Participants younger than 45 years old (OR = 2.3 and 95% CI = 1.6-3.4 for themselves; OR = 2.2 and 95% CI = 1.4-3.3 for their daughters) were more likely to accept the vaccination than those aged 45 years old and above. CONCLUSION: Knowledge about HPV and the HPV vaccine is generally poor in Thai women. However, the acceptability of the HPV vaccine is good. Knowing about HPV and age under 45 years predict the acceptability of the HPV vaccine.

Targeted delivery of small interfering RNA using reconstituted high-density lipoprotein nanoparticles.

RNA interference holds tremendous potential as a therapeutic approach, especially in the treatment of malignant tumors. However, efficient and biocompatible delivery methods are needed for systemic delivery of small interfering RNA (siRNA). To maintain a high level of growth, tumor cells scavenge high-density lipoprotein (HDL) particles by overexpressing its receptor: scavenger receptor type B1 (SR-B1). In this study, we exploited this cellular characteristic to achieve efficient siRNA delivery and established a novel formulation of siRNA by incorporating it into reconstituted HDL (rHDL) nanoparticles. Here, we demonstrate that rHDL nanoparticles facilitate highly efficient systemic delivery of siRNA in vivo, mediated by the SR-B1. Moreover, in therapeutic proof-of-concept studies, these nanoparticles were effective in silencing the expression of two proteins that are key to cancer growth and metastasis (signal transducer and activator of transcription 3 and focal adhesion kinase) in orthotopic mouse models of ovarian and colorectal cancer. These data indicate that an rHDL nanoparticle is a novel and highly efficient siRNA carrier, and therefore, this novel technology could serve as the foundation for new cancer therapeutic approaches.

Comparison of outcomes for patients with cervical cancer who developed bone metastasis after the primary treatment with concurrent chemoradiation versus radiation therapy alone.

OBJECTIVES: The aims of this study were to retrospectively compare outcomes for patients with cervical cancer who developed bone metastasis later after the primary treatment at the time of diagnosis of cervical cancer with concurrent chemoradiation (CCRT) to radiation therapy alone (RT). METHODS: We retrospectively analyzed the patients with cervical cancer during the period from January 1998 to December 2007. Of these, 11 patients who received CCRT and 24 patients who received RT went on to develop bone metastasis. RESULTS: Among 4620 patients with cervical cancer, 51 patients had bone metastases. Sixteen patients were excluded including 10 patients with unavailable records and 6 patients who did not receive CCRT or RT at the time of diagnosis of cervical cancer. Thirty-five patients who had bone metastasis received primary treatment with CCRT or RT. The 2 groups of patients (CCRT vs RT) were similar in age, histologic cell type, and the International Federation of Gynecology and Obstetrics stages. The characteristics of bone metastasis in both groups were also not significantly different. The patients who received CCRT did not have a better overall survival than the patients who received RT (median, 19 vs 22 months; 95% confidence interval [CI], 14.68-23.32 vs 8.56-35.44). They were comparable in the interval from cervical cancer diagnoses to diagnoses of bone metastasis (CCRT group: median, 14 months; 95% CI, 9.14-18.86; RT group: median; 15 months; 95% CI, 10.20-19.80) and the survival after diagnosis of bone metastasis between both groups (CCRT group: median, 4 months; 95% CI, 0.76-7.24; RT group: median, 7 months; 95% CI, 4.70-9.30). CONCLUSIONS: Our retrospective analysis showed that there were no differences in survival benefits observed between the patients with cervical cancer who developed bone metastases later after the primary treatment with CCRT and RT.

Female hospital-based healthcare professionals' knowledge of cervical cancer, HPV and attitudes towards HPV vaccination.

OBJECTIVE: To find out female hospital-based healthcare professionals' knowledge of cervical cancer, HPV and attitudes towards HPV vaccination. DESIGN: A descriptive cross-sectional hospital-based study. SETTING: Ramathibodi Hospital, Mahidol University, Bangkok, Thailand. SAMPLE: A total of 350 female hospital-based healthcare professionals who had not been diagnosed as having any cancer and willing to participate in the study. METHODS: Participants completed written consent and an anonymous questionnaire and knowledge of cervical cancer, HPV and attitudes towards HPV vaccination were the main outcome measures. RESULTS: Among 300 responders, the mean age was 36.1 years. Most of them were married with children and had received university education. Nursing assistants accounted for 47.1%, and their income per month was about 5,001-15,000 baht. Most (56.3%) had only one lifetime sexual partner. Sixty-eight to 85.3% have a good knowledge of cervical cancer and Pap smear. However, only 12.0 to 58.3% have some knowledge of HPV, and less than fifty percent of them have knowledge of HPV vaccination. Nevertheless, 51.7 to 60.7% of them have good attitudes toward vaccination. Their age and income might influence their attitudes about having themselves vaccinated, and their career might be a factor which altered their attitudes about having their daughter vaccinated if they have one. CONCLUSIONS: Female hospital-based healthcare professionals have a good knowledge about cervical cancer and Pap smears, but they need motivation to have Pap tests regularly. More information regarding HPV and vaccination is needed to provide to them for cervical cancer prevention and best practices.

Bone metastasis in cervical cancer patients over a 10-year period.

INTRODUCTION: Cervical cancer is the major cancer burden in developing countries. Bone is the third most common site of distant metastasis after the lungs and liver. Therefore, the aims of this study were to find the incidence and clinical characteristics of bone metastasis in our hospital. PATIENTS AND METHODS: Fifty-one cervical cancer patients with bone metastasis during the period from January 1998 to December 2007 were recruited. All patients' medical records were reviewed and analyzed. RESULTS: Among 4620 cervical cancer patients, there were 51 patients (1.1%) who had bone metastases. Ten patients' medical records were not found; thus, 41 patients were available for evaluation. The median age of the patients was 49 years. International Federation of Gynecology and Obstetrics stage IIB was the most common stage (43.9%). Most patients had squamous cell carcinoma (80.48%) and received radiation therapy alone as their primary treatment (58.53%). The most common presenting symptom was pain (78.04%). Most of the patients had multiple bone lesions and extrapelvic bone metastases. The lumbar spine was the most common site (36.36%). Sixteen patients (39.02%) were treated by palliative radiation therapy. The median overall survival was 23 months. CONCLUSIONS: Bone metastases could be found at all stages. Common sites were the bone beyond the radiation field of their primary treatment. It was found at a median of 16 months after cervical cancer diagnosis. Currently, there are many varieties of treatment that result only in palliation. This group of patients has a poor prognosis.

Can vaginal misoprostol effectively increase rate of a satisfactory colposcopy? A randomized double-blind placebo-controlled trial.

OBJECTIVE: To evaluate the effectiveness of vaginal misoprostol in overcoming an unsatisfactory colposcopy in the patients who had abnormal cervical cytology and to evaluate side effects of vaginal misoprostol. METHODS: Sixty patients with an unsatisfactory colposcopy during the period of September 2007-November 2008 were recruited and randomly allocated to receive either two tablets of 200 microg misoprostol (400 microg) or two tablets of similar-looking placebo vaginally. Colposcopic re-examination was performed approximately 6 h later. The results and side effects before and 2 weeks after the colposcopic re-examination were recorded. RESULTS: Six out of 30 patients in the misoprostol group (20.0%) had a satisfactory colposcopic re-examination compared with 2 out of 27 patients (7.4%) in the placebo group without statistically significant difference (P = 0.172). Three patients in the placebo group dropped out due to not present at the appointment time. Six out of 30 patients (20.0%) and 1 out of 30 patients (3.3%) in the misoprostol group had side effects before and 2 weeks after the colposcopic re-examination orderly. Twenty-seven patients in the placebo group did not have any side effects before and 2 weeks after the colposcopic re-examination. All side effects occurred were minimal and well tolerated. CONCLUSIONS: Four hundred micrograms of vaginal misoprostol were not proved to be effective in converting an unsatisfactory to a satisfactory colposcopy.