Bone Metastatic Cancer Patients Research Articles

Bone targeted miRNA delivery system for miR-34a with enhanced anti-tumor efficacy to bone-associated metastatic breast cancer

Bone metastatic cancer is the most common occurrence in breast cancer, and the treatment is also facing great challenges. MicroRNA-34a (miRNA-34a) is a promising anti-cancer miRNA for gene therapy to bone metastatic cancer patients. However, the lack of specificity to bone and low accumulation at the site of bone tumor remains the major challenge when used bone-associated tumor. To solve this problem, a bone-targeted vector for delivery of miR-34a to bone metastatic breast cancer was constructed by using the commonly used gene vector branched polyethylenimine 25 k (BPEI 25 k) as the skeleton and linking with alendronate (ALN) moieties for bone targeting group. The constructed gene delivery system PCA/miR-34a can efficiently prevent miR-34a from degradation during blood circulation and enhance the specific bone delivery and distribution. PCA/miR-34a nanoparticles can be uptake into tumor cells through clathrin and caveolae-mediated endocytosis, and directly regulate the expression of oncogenes, thus promoting tumor cell apoptosis and relieving bone tissue erosion. The results of experiments in vitro and in vivo confirmed that the constructed bone-targeted miRNA delivery system PCA/miR-34a can enhance the anti-tumor efficacy in bone metastatic cancer, and provide a potential strategy for gene therapy in bone metastatic cancer.

Predicting Response to Radiotherapy in Breast Cancer-Induced Bone Pain: Relationship Between Pain and Serum Cytokine Expression Levels After Radiotherapy.

Breast cancer is the second leading cause of cancer-associated death in women. Herein, we explored the associations of cytokines, pain, and bone metastasis between patients before and after radiotherapy in breast cancer with bone metastasis. The pain caused by metastasis was effectively relieved by external radiation therapy. However, the underlying mechanisms remain unknown. In this case-controlled study, we enrolled healthy individuals (n = 10) and bone metastatic cancer patients (n = 30). Peripheral venous blood samples were collected from healthy controls, and one week before and after radiotherapy, the peripheral venous blood and clinical characteristics of cancer patients were collected. We analyzed the blood cytokine profile, quality of life (QOL), and pain score of patients pre- and post-radiotherapy to explore the possible causes of pain relief. Both the pain score and QOL significantly improved after radiotherapy. The serum cytokine profiles of patients were significantly different before radiotherapy than after. Meanwhile, only three cytokines differed between post-radiotherapy and healthy controls. We believe radiotherapy stimulated local immune storms in bone tissue and promoted significant changes in cytokines pre- and post-radiotherapy. Therefore, the bone microenvironment of early breast cancer patients with bone metastasis pain can be restored after radiotherapy. Restoring a healthy bone environment can not only relieve the pain but also improve the patient's QOL. Tumor cells can effectively activate immune cells in the bone microenvironment through direct interaction, releasing many factors and promoting bone metastasis. Early local radiotherapy of bone metastases can restore the microenvironment and improve the QOL and prognosis of patients, thereby comprehending a novel target for prevention, treatment, and therapy of bone metastases.

One changing and challenging scenario: the treatment of cancer patients with bone metastases by bisphosphonates and denosumab, the cost-benefit evaluation of different options, and the risk of medication-related osteonecrosis of the jaw (MRONJ).

Antiresorptive drugs (bisphosphonates and denosumab) have become the cornerstone of medical supportive treatment of bone metastases in solid cancer patients. In the beginning, the choice of available antiresorptive agents was limited to bisphosphonates and the treatment options restricted principally to monthly pamidronate and monthly zoledronic acid. Introduction of new antiresorptive therapies (monthly denosumab) and schedules (zoledronic acid every 3 months, upfront or after initial period of monthly infusion) in the last decade increased the range of available options, thus challenging treatment decision making. Direct and indirect costs of very different treatment options are difficult to interpret in a global cost–benefit analysis. In addition, awareness of the increased risk of medication-related osteonecrosis of the jaw (MRONJ) in bone metastatic cancer patients receiving long-term antiresorptive medications is likely to influence therapy choice in the real-life scenario. We discuss the possible threat of MRONJ risk underestimation and the need for long-term risk stratification of patients based on actuarial data, the role of bisphosphonates and denosumab in that scenario, and the emerging role of surgical therapy to successfully cure MRONJ, in the light of the improved quality of life and survival of patients with bone metastases from solid cancers.

MicroRNA Prognostic Signature for Postoperative Success of Metastatic Orthopedic Cancers: Implications for Precision Microsurgery.

The importance of miRNA prognostic signature in cancer, particular cancer metastasis is increasingly being realized. Bone metastasis from several primary human cancers can be managed in clinics by surgical intervention but the prognostic impact of miRNA signature on post-surgery outcome of patients is unknown. This study evaluated a miRNA signature for post-operative outcome of patients with bone metastatic disease. First, the miRNAs, miR-135, miR-203, miR-10b, miR-194, miR-886, and miR-124 were evaluated in bone metastatic tissues, relative to adjacent control tissue. The cohorts of samples (n = 44) consisted of bone metastatic cancer patients with primary lung (n = 18) or breast cancer (n = 26). miR-203 was significantly down-regulated while miR-10b was significantly up-regulated in bone metastasis. Additionally, miR-135 was significantly differentially expressed in the primary lung cancer patients while miR-194 in primary breast cancer patients. The low miR-203- high miR-10b expression was designated high risk group and, compared to the low risk group (high miR-203-low miR-10b expression). Patients with the signature high risk fared significantly better with surgical intervention, in terms of survival at 12 months time point (40% survival with surgery vs. 10% survival without surgery), as revealed by retrospective analysis of patient data. This work reveals potential utilization of miRNA expression levels in not only the general prognosis of cancer metastasis but also the prognosis of surgical intervention with implication for better stratification of patients.

Improved Radium-223 Therapy with Combination Epithelial Sodium Channel Blockade.

Background: Radium-223 dichloride ([223Ra]RaCl2) is the first approved alpha particle-emitting therapy and is indicated for treatment of bone metastatic castrate resistant prostate cancer. Approximately half of the dose is absorbed into the gastrointestinal (GI) tract within minutes of administration, limiting disease-site uptake and contributing to toxicity. Here, we investigate the role of enteric ion channels and their modulation for improved therapeutic efficacy and reduced side effects. Methods: Utilizing primary human duodenal organoids (enteroids) as in vitro models of the functional GI epithelium, we found that Amiloride (ENaC blocker) and NS-1619 (K+ channel activator) presented significant effects in 223Ra membranal transport. The radioactive drug distribution was evaluated for lead combinations in vivo, and in osteosarcoma and prostate cancer models. Results: Amiloride shifted 223Ra uptake in vivo from the gut, to nearly double the uptake at sites of bone remodeling. Bone tumor growth inhibition with the combination as measured by bioluminescent and X-ray imaging was significantly greater than single agents alone, and the combination resulted in no weight loss. Conclusion: This combination of approved agents may be readily implemented as a clinical approach to improve outcomes of bone metastatic cancer patients with the benefit of ameliorated tolerability.

Measurement of Bone Metastatic Tumor Growth by a Tibial Tumorigenesis Assay.

Bone metastasis is a frequent and lethal complication of many cancer types (i.e., prostate cancer, breast cancer, and multiple myeloma), and a cure for bone metastasis remains elusive. To recapitulate the process of bone metastasis and understand how cancer cells metastasize to bone, intracardiac injection and intracaudal arterial animal models were developed. The intratibial injection animal model was established to investigate the communication between cancer cells and the bone microenvironment and to mimic the setting of prostate cancer patients with bone metastasis. Given that detailed protocols of intratibial injection and its quantitative analysis are still insufficient, in this protocol, we provide hands-on procedures for how to prepare cells, perform the tibial injection, monitor tibial tumor growth, and quantitatively evaluate the tibial tumors in pathological samples. This manuscript provides a ready-to-use experimental protocol for investigating cancer cell behaviors in bone and developing novel therapeutic strategies for bone metastatic cancer patients.

Single-photon Emission CT Combined with Spiral CT for Early Detection and Localization of Bone Metastasis: A Review.

Bone metastasis is common in cancer. Evaluating the metastatic status in cancer is of utmost importance in order to provide the best patient's management. Bone scintigraphy is widely used for evaluation of bone metastasis. It has high sensitivity with limited specificity. Planar bone scintigraphy has been shown to have increased radiotracer uptake without accurate anatomic localization and characterization. Hybrid Single-Photon Emission Computed Tomography/Computerized Tomography (SPECT/CT) system has been developed by combination of SPECT and CT. Accurate lesion localization and discrimination of equivocal bone lesions is an advantage in this hybrid technique. It improves diagnostic accuracy by differentiation of benign bone lesions from malignant ones due to their morphological changes. So, SPECT/CT improves the specificity of bone scintigraphy leading to better outcomes in diagnosis and treatment outcomes of bone metastatic cancer patients. In here, we discussed the prognostic value of bone scintigraphy and SPECT/CT in bone metastasis with our clinical experience and review of the literature.

Long-term outcomes of prolonged bisphosphonates more than 2 years in bone metastatic breast cancer: risk vs benefit.

Bisphosphonates are the mainstay therapeutic options for prevention of skeletal-related events and generally used for up to 2 years in bone metastatic cancer patients. We aimed to evaluate the long-term outcomes of prolonged (> 2 years) bisphosphonate usage in bone metastatic breast cancer (BMBC) patients. Ninety-nine BMBC patients who had prolonged bisphosphonates were evaluated retrospectively for long-term outcomes and survival rates. Median duration of bisphosphonate therapy was 46.8 (24-198) months. Seven patients had bisphosphonate-related adverse events (osteonecrosis of the jaw (ONJ) (n = 6), ONJ and renal failure (n = 1)). Bisphosphonate was switched to another one because of bone metastasis progression in more than one-third of the patients (n = 36, 36.3%). The patients who had bisphosphonate switch therapy had statistically significant longer overall survival (p < 0.01). Neither duration nor type of bisphosphonates had effect on frequency of bisphosphonate-related adverse events. Bisphosphonates might be prolonged for more than 2 years in BMBC patients with an acceptable toxicity profile. In addition, bisphosphonates switch therapy should be preferred in those with progressive bone metastasis since it might contribute to better survival despite bisphosphonates could not have been shown to have survival benefit in previous studies.

MTOR inhibitor and bone-targeted drugs break the vicious cycle between clear-cell renal carcinoma and osteoclasts in an in vitro co-culture model

The skeleton is one of the most common sites of metastatic spread from advanced clear-cell renal carcinoma (ccRCC). Most of the bone lesions observed in RCC patients are classified as osteolytic, causing severe pain and morbidity due to pathological bone destruction. Nowadays, it is well known that cancer induced bone loss in lytic metastasis is caused by the triggering of a vicious cycle between cancer and bone resident cells that leads to an imbalance between bone formation and degradation. Targeting the mammalian target of rapamycin (mTOR) is an efficient treatment option for metastatic renal carcinoma patients. Moreover, bone targeted therapy could benefit bone metastatic cancer patients caused by advanced RCC. However, more data is needed to support the hypothesis of the beneficial effect of a combined therapy. The aim of this work is to investigate the effect of targeting mTOR and the sequential combination with bone targeted therapy as a strategy to break the vicious cycle between ccRCC cells and osteoclasts. A previously optimized fully human co-culture model is used to mimic the crosstalk between Caki-2 cells (ccRCC) and osteoclasts. Cells are treated at fixed timing with everolimus, zoledronic acid and denosumab as single or sequential combined treatment. We show that Caki-2 cells can induce osteoclast cells differentiation from isolated human monocytes, as demonstrated by specific tartrate-resistant acid phosphatase (TRAP) staining and f-actin ring formation, in a statistically significant manner. Moreover, differentiated osteoclasts proved to be functionally active by pit formation assay. Caki-2 cells co-cultured with osteoclasts acquire a more aggressive phenotype based on gene expression analysis. Interestingly, the sequential combined treatment of everolimus and zoledronic acid is the most effective in the inhibition of both Caki-2 cells survival and osteoclastogenic potential, making it an effective strategy to inhibit the vicious cycle of bone metastasis. At preclinical level, this observation confirms the value of our co-culture model as a useful tool to mimic the bone microenvironment and to assess drug sensitivity in vitro. A better understanding of the molecular mechanisms involved in tumor-bone cells crosstalk will be investigated next.

Evaluation the quality of life in patients with bone metastases: evidence-based update

There is increasing recognition of the importance of the quality of life (QOL) since it became an endpoint of clinical practices, a variety of disease-specific quality of life instruments have been developed and used as a means of assessing clinical interventions. Traditional evaluation was performed by researchers who generally ignore the subjective feeling from patiens. At present, many evaluation criteria has paid more attention to the patient's subjective feelings into the scoring system, the assessment tool of patient-reported outcome (PRO) is usually the patient self-rating scale or questionnaire. As a common complications of terminal stages of cancer, the treatment of bone metastases is usually conservative, improving or maintaining the quality of life of patients has been the consensus. Bone metastases trials have employed QLQ-C30 which raised by Europe Organization for Research and Treatment of Cancer (EORTC) or other generic health related QOL measurement, such as functional assessment of cancer therapy-general (FACT-G) and the MOS 36-item short from health survey (SF-36). But they lack specificity and accuracy in the assessment. EORTC QLQ-BM22 scale is considered as a bone metastases specific scale which was introducted in 2009, after the reliability, validity, cross-cultural adaptability verification, with the combination of EORTC QLQ-C30 or used alone, has increasingly applied to clinical trials and practice for QOL assessment of bone metastatic cancer patients in recent years. It complete the EORTC evaluation system, which has significant value for assessing efficacy. EORTC QLQ-BM22 has been translated into simplified Chinese and evaluated its utility, could be used in China as an effective questionnaire in evaluating the QOL for patients with bone metastases. In this study, current situation and progress of QOL measuring tools for bone metastases patients is reviewed, in order to help researchers choose appropriate scale and better assess the efficacy, find the pros and cons to guide the clinical treatment decision.

Arrhythmias during and after zoledronic acid infusion patients with bone metastasis.

e20614 Background: Zoledronic acid (ZA) is one of the important biphosphanate which is widely used in bone metastatic cancer patients and osteoporotic patients. In a few studies it has been reported that treatment with biphosphonate was associated with an increased risk for atrial fibrillation. We aim to evaluate the arrhythmias that developed during infusion and immediately after infusion of the ZA. Methods: Bone metastatic 52 patients were included in study group. All of patients had totally 24 hour holter motorization during the first dose ZA infusion day. All of patients had 4 hours basal cardiac rhythm records before ZA infusion and about 19 hours after infusion. A short survey including demographic data and past medical history was completed. None of patients had clinically important arrhythmias before ZA infusion. We divided arrhythmias as supraventricular and ventricular. We evaluated arrhythmias pre, during and after ZA infusion. ZA were administered 4 mg IV in 15 minutes. Results: Thirty three of patients (63,5%) were male and 19 (36,5%) female. Median age of the patients was 52 (27-73). Most frequent cancers were breast (25%) and lung cancer (15,3%).Twelve (23%) patients had history of mediastinal radiotherapy. Three patients had history of myocardial infarction. In basal records we detected that twenty four of patients had supraventricular tachycardia (SVT) or ventricular premature systole (VPS). Fifteen of patients had SVT and fourteen had VPS during infusion period. After infusion period 48 of patients had SVT and 41 had VPS. Only 3 patients had none of arrhythmia after infusion. Three of the patients had sinusoidal arrhythmia and two had Mobitz type 2 bloc after infusion and also one of our patient who had no history of co morbidities and had SVT basal records, developed atrial fibrillation that refractory to medical cardioversion after 10 days of seventh dose ZA infusion. Conclusions: In this study we found that both SVT and VPS increased in cancer patients treated with ZA. Furthermore ZA may induce clinically important arrhythmias.

Samarium-153 effectiveness in the quality of life of bone metastatic cancer patients.

e20635 Background: Osseous metastases in patients suffering from cancer, is a multifactor symptom which affects quality of life. Methods: The Samarium group (SG) n=53 patients received a single bolus infusion of (153)Sm (37 MBq/kg), and we compared with a Control group(CG) n=37. Both groups of patients who had metastatic bone cancer requiring analgesia answered the following questionnaires: Greek Brief Pain Inventory (GBPI), Brief Multidimensional Life Satisfaction Scale (BMLSS), Hospital Anxiety Depression Scale (HADS), ECOG/WHO/Zubrod score and Hospital Satisfaction Scale (HSS),day 0 and 30±3 days after the intervention start.There was no statistically significant(SS) difference for the variables(V-) sex, age, pain’s localization and lodging. Results: Pain severity and pain interference change over time was the same for both groups P= 0.0005. For the V- HADS-anxiety, SG showed no SS improvementP= 0.397, something which was achieved for the CG,P= 0.031. The V- HADS improved SS for both groups P= 0.031 and P= 0.003 respectively which changed over time with the same way. This may show the strong relation between pain and depression. The V- ECOG also changed over time in the same way for both SG(P=0.005) and CG (P=0.014). The percentage of patients who changed scale towards improvement for their quality of life was 24,3% for the CG and 15,1% for the SG. The V-BMLSS change over time was similar for both groups. There was no SS difference for the V-HSS towards medical and nursing personnel for both groups (P=1.000). Myelotoxicity for the SG was moderate and reversible, as well as the side effects for the CG. Finally an equivalence test between the two groups, showing that both treatment regimens were equivalent for all variables: Pain severity=0.034, Pain interference=0.009, HAD-anxiety P=0.048, HAD-depression=0.033, BMLSS=0.016, ECOG=0.0005. During our study among the patients of the SG in some cases the stabilization of bone lesions was observed, and in one case partial and/or complete remission. Conclusions: Administration of (153)Sm is another equivalent option for those patients who are intolerant or resistant to medication treatment having a better cost-effectiveness result.

Efficacy of electroacupuncture for bone metastatic cancer patients with neuropathic pain: A randomized controlled trial

9534 Background: Bone metastatic lung cancer or breast cancer pts often have neuropathic pain which has poor reaction to opioids. A clinical trial is to investigate efficacy of electroacupuncture (EA) in cancer pts with neuropathic pain. Methods: Inclusion criteria were: bone metastatic lung cancer or breast cancer pts at least one of the symptoms as burning pain, shooting pain, dysesthesias, or allodynia; numerical rating scale (NRS) of pain intensity &gt;=4.0; stable dose of opioids for at least 72 hours. Exclusion criteria were: chemotherapy or radiotherapy from 15 days before or during the study. Hormone or adjuvant therapy could be started before the study, but the dose could not be changed afterwards. Opioids could be increased step by step according to the pain. EA was manipulated with low-frequency (2 Hz) and high-frequency (100 Hz) dense-disperse waves alternatively, 30 minutes per day, 5-day therapy with 2-day interval, 3 weeks continuously. Acupoints were selected as ST36, SJ3, SJ5, LI3, LI4 bilateral. Controlled group accepted non-acupoint treatment close the right acupoint. Results: Ninety pts with neuropathic pain participated in a randomized, controlled design trial from 2006 to 2008. Overall, 45 pts accepted EA treatment and 34 completed the study. 45 pts accepted controlled and 32 completed. There was no difference of initial NRS which was 6.3±2.1 in EA group and 7.2±1.8 in controlled, p = 0.0669. Analysis showed a significant difference of NRS between EA group (3.2±1.7) and controlled (5.4±2.2) at the end of survey, p &lt; 0.0001. The decrease in NRS was significantly higher in the EA group compared to the controlled (3.2±1.7 vs. baseline, p &lt; 0.0001; 5.4±2.2 vs. baseline, p = 0.0007). Mean analgesics consumption variety in EA group was as follows: transdermal fentanyl increased by 51.2% (16 pts) and 77.3% (21 pts) in EA and controlled, p = 0.0005. Oxycodone increased by 85.5% (18 pts) and 115.6% (11 pts) in EA and controlled, p = 0.0183. Functional Assessment of Cancer Therapy-General (FACT-G) scale had no difference at the beginning, while it were 102±18 and 85±12 in EA group and controlled at the end of survey, p &lt; 0.0001. Conclusions: EA can alleviate neuropathic pain of bone metastatic cancer pts, decrease the analgesics consumption, and promoting quality of life. No significant financial relationships to disclose.

Pretreatment serum level of HER2/nue as a prognostic factor in metastatic prostate cancer patients about to undergo endocrine therapy

Overexpression of the HER2 receptor protein and amplification of the HER2 gene has been implicated in tumor development and progression, and has been associated with a poor prognosis in several types of cancer. The aim of this study was to evaluate whether pretreatment serum HER2 levels can be used to predict biochemical recurrence-free survival in prostate cancer patients about to undergo endocrine therapy. The study population consisted of 379 untreated patients with histologically diagnosed prostate cancer: 197 with T2N0M0, 93 with T3N0M0, 19 with TxN1Mx, and 70 with TxNxM1. Serum HER2 levels were assessed in the prostate cancer patients prior to treatment as well as in a control group of 100 patients with histologically confirmed non-cancer. Biochemical recurrence-free curves for the patients were investigated separately using the Kaplan-Meier method. The mean level of HER2 in serum was significantly higher in prostate cancer patients than non-prostate cancer patients (P = 0.006). Also, the serum HER2 level was significantly higher in bone metastatic cancer patients (14.3 +/- 6.3 ng/mL) than in non-metastatic patients (T2: 11.9 +/- 2.3 ng/mL, P = 0.003; T3: 12.2 +/- 2.8 ng/mL, P = 0.011). The metastatic patients were divided into those with low and high HER2 levels using a cutoff value of 12.6 ng/mL based on receiver-operating characteristic curves. The biochemical recurrence-free rate was significantly poorer in patients with a high HER2 level (P = 0.0078, log-rank test). Multivariate Cox logistic regression analysis demonstrated that the pretreatment serum HER2 value (P = 0.022), serum prostate-specific antigen value (P = 0.018), and extent of disease score (P = 0.027) were independent predictors of recurrence. The pretreatment serum HER2 level may be a useful independent prognostic factor that is associated with a high risk of biochemical recurrence in metastatic prostate cancer patients about to undergo endocrine therapy.

The circadian rhythm of biochemical markers of bone resorption is normally synchronized in breast cancer patients with bone lytic metastases independently of tumor load

Background Bone metastases are devastating events resulting in disruption of local bone remodeling processes. Physiological bone turnover has a circadian rhythm. No data are available on the circadian pattern of bone turnover markers in patients with bone metastases. Methods Twenty post-menopausal women with breast cancer (BC) at first disease relapse and at least one bone metastasis were consecutively recruited. Twenty healthy women served as controls. Patients were free from concomitant chemotherapy/endocrine therapy. Throughout a 24-h period, urine samples were collected at 4-h intervals, and blood samples were collected at 4-h intervals between 08:00 and 24:00, and at 2-h intervals between 24:00 and 08:00. Serum osteocalcin (OC), total and bone-alkaline phosphatase (tALP and bALP, respectively) and C-terminal telopeptide of type I collagen (CTX), and urinary NTX and free deoxypyridinoline (fDPD) were measured together with serum parathyroid hormone (PTH) and serum and urinary calcium and phosphorus. Temporal variations of measured analytes were assessed by ANOVA and the COSINOR model. Results At 08:00, patients had higher levels of bone resorption indices (NTX, CTX and fDPD) than controls ( p < 0.0001). tALP and bALP, but not OC, were higher in patients than controls ( p < 0.001). PTH, serum and urinary calcium and urinary phosphorus did not differ between groups; serum phosphorus was higher in controls ( p < 0.0001). A circadian rhythm was evident for CTX and fDPD values in both patients and controls. A circadian rhythm in NTX, OC, phosphorus and PTH was apparent in controls only. However, it was detected also in patients when percent changes from MESOR were considered. Serum phosphorus showed a circadian rhythm, while no rhythm was detected for tALP, bALP, serum and urinary calcium. The rhythmicities in cancer patients were normally synchronized, and rhythmic parameters were independent of tumor load in the skeleton, age and menopausal status. Conclusions This is the first study to yield information on the maintenance of the temporal program of bone turnover in bone metastatic cancer patients. Whether administration of bisphosphonates in the nighttime leads to a different outcome with respect to the current administration in the morning is a matter of future research.

Zoledronic Acid-Related Angiogenesis Modifications and Survival in Advanced Breast Cancer Patients

The proven antiangiogenic activity of zoledronic acid, a third-generation bisphosphonate widely used in bone metastatic cancer patients, led us to investigate if the vascular endothelial growth factor (VEGF)-related zoledronic acid modifications are correlated with survival advantages in advanced breast cancer patients. Forty-two consecutive breast cancer patients with scintigraphic and radiographic evidence of bone metastases were treated with a single infusion of 4 mg zoledronic acid before anticancer chemotherapy. The patients were prospectively evaluated for circulating levels of VEGF and interferon-gamma (IFN-gamma) just before and at 1, 2, 7, and 21 days after zoledronic acid infusion. Afterward, clinical outcome was prospectively monitored. The basal serum VEGF median levels were significantly decreased at each time point, but the major reduction was recorded 21 days after the infusion. In particular, 25 patients of 42 (59.5%) experienced a reduction of at least 25% in the VEGF circulating levels. In contrast, no statistically significant modifications of the IFN-gamma serum levels were recorded. We stratified patients on the basis of this VEGF reduction 21 days after the infusion. No differences in patient features were recorded between those with or without the VEGF reduction. The analysis of survival showed that patients with a reduction in the VEGF circulating levels had a longer time to first skeletal-related event (p = 0.0002), time to bone progression disease (p = 0.0024), and time to performance status worsening (p = 0.0352) than those without the VEGF reduction. No statistically significant differences were recorded in terms of overall survival and time to visceral progression. This study confirms that zoledronic acid could have an in vivo antiangiogenic property and that the VEGF modifications may represent a surrogate marker able to predict time to first skeletal-related event, time to bone progression disease, and time to worsening of performance status.

Determination and biological relevance of serum cross-linked type I collagen N-telopeptide and bone-specific alkaline phosphatase in breast metastatic cancer

Bone metastasis is a frequent complication of cancer disease. The metastatic spread of cancer to bone is common to many different malignancies, particularly breast (ca. 73%), prostate (ca. 68%) and lung (ca. 36%) cancers. Metastases to bone cause increased bone resorption both from direct effects of the tumor itself and thought osteoclastic activation. The diagnosis and follow-up of bone metastatic cancer patients usually relies on skeletal X-ray and bone scintigraphy. However, the development of biochemical markers, used as indicators of bone metabolism, provides data useful in the clinical practice. The most important markers for bone remodeling process, bone formation and resorption, are bone-specific alkaline phosphatase (BAP) and N-telopeptide of type I collagen (NTx), respectively. In this report, we applied two solid-phase immunoassays used for the determination of BAP and NTx in serum of breast cancer (BC) post-menopausal women with bone metastasis and healthy individuals. BAP level in patients was found to be 45.72±12.92 U/l, while the normal range for healthy individuals was 14.2–42.7 U/l. The respective level of serum NTx was 19.20±8.87 nM bone collagen equivalents (BCE) for patients and 15.9±3.8 nM BCE for healthy women. Correlation of the obtained data showed elevated levels for both markers indicating high rate of bone degradation in breast metastatic cancer.