Curcumin-Induced Apoptosis in Human Hepatocellular Carcinoma J5 Cells: Critical Role of Ca+2-Dependent Pathway

The antitumor effects of curcumin, a natural biologically active compound extracted from rhizomes of Curcuma longa, have been studied in many cancer cell types including human hepatocellular carcinoma (HCC). Here, we investigated the effects of Ca2+ on curcumin-induced apoptosis in human HCC J5 cells. The abrogation of mitochondrial membrane potential (ΔΨm), the increase of reactive oxygen species (ROS) production, and calcium release were demonstrated with flow cytometry as early as 15 minutes after curcumin treatment. In addition, an increase level of cytochrome c in the cytoplasm which led to DNA fragmentation was observed. To verify the role of Ca2+ in curcumin-induced apoptosis, 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), an intracellular calcium chelator, was applied. Cell viability was increased, but ΔΨm, ROS production, activation of caspase 3, and cell death were decreased in J5 cells pretreated with BAPTA for 2 h followed by the treatment of 25 μM curcumin. These results suggest that the curcumin-induced apoptosis in human HCC J5 cells is via mitochondria-dependent pathway and is closely related to the level of intracellular accumulation of calcium

Role of p21WAF1 and p27KIP1 in Predicting Biochemical Recurrence for Organ-confined Prostate Adenocarcinoma

Both p21WAF1 and p27KIP1 have been reported as prognostic markers predicting biochemical failure for prostate cancers. We examined the expression and prognostic significance of p21WAF1 and p27KIP1 in organ-confined (pT2) prostate cancer patients. Methods: The medical records of 53 pT2 prostate adenocarcinomas were analyzed retrospectively. Radical prostatectomy specimens were stained using anti-p21WAF1 and anti-p27KIP1 antibodies. Biochemical relapse was defined as 2 consecutive elevations in serum prostate specific antigen (PSA) level > 0.2 ng/mL with an interval of more than 3 months. The prognostic significance of p21WAF1 and p27KIP1 expression was assessed. Results: p21WAF1 immunoreactivity was found in 19 patients (35.8%). Twenty-nine tumors (54.7%) had decreased p27KIP1 expression. Both markers were not associated with Gleason scores (p = 1.00 for both). At a median follow-up of 49 months, 15 patients (28.3%) experienced biochemical recurrence. Both p21 and p27 had no prognostic significance in log-rank test (p = 0.98 and p = 0.64, respectively). Conclusion: p21WAF1 and p27KIP1 expression have no role in predicting biochemical relapse for stage pT2 prostate cancers

Tumor burden talks in cancer treatment with PEGylated liposomal drugs.

PURPOSE: PEGylated liposomes are important drug carriers that can passively target tumor by enhanced permeability and retention (EPR) effect in neoplasm lesions. This study demonstrated that tumor burden determines the tumor uptake, and also the tumor response, in cancer treatment with PEGylated liposomal drugs in a C26/tk-luc colon carcinoma-bearing mouse model. METHODS: Empty PEGylated liposomes (NanoX) and those encapsulated with VNB (NanoVNB) were labeled with In-111 to obtain InNanoX and InVNBL in high labeling yield and radiochemical purity (all >90%). BALB/c mice bearing either small (58.4±8.0 mm(3)) or large (102.4±22.0 mm(3)) C26/tk-luc tumors in the right dorsal flank were intravenously administered with NanoVNB, InNanoX, InVNBL, or NanoX as a control, every 7 days for 3 times. The therapeutic efficacy was evaluated by body weight loss, tumor growth inhibition (using calipers and bioluminescence imaging) and survival fraction. The scintigraphic imaging of tumor mouse was performed during and after treatment. RESULTS: The biodistribution study of InVNBL revealed a clear inverse correlation (r (2) = 0.9336) between the tumor uptake and the tumor mass ranged from 27.6 to 623.9 mg. All three liposomal drugs showed better therapeutic efficacy in small-tumor mice than in large-tumor mice. Tumor-bearing mice treated with InVNBL (a combination drug) showed the highest tumor growth inhibition rate and survival fraction compared to those treated with NanoVNB (chemodrug only) and InNanoX (radionuclide only). Specific tumor targeting and significantly increased tumor uptake after periodical treatment with InVNBL were evidenced by scintigraphic imaging, especially in mice bearing small tumors. CONCLUSION: The significant differences in the outcomes of cancer treatment and molecular imaging between animals bearing small and large tumors revealed that tumor burden is a critical and discriminative factor in cancer therapy using PEGylated liposomal drugs

Mirtazapine Inhibits Tumor Growth via Immune Response and Serotonergic System

<div><p>To study the tumor inhibition effect of mirtazapine, a drug for patients with depression, CT26/<em>luc</em> colon carcinoma-bearing animal model was used. BALB/c mice were randomly divided into six groups: two groups without tumors, i.e. <em>wild-type</em> (no drug) and <em>drug</em> (mirtazapine), and four groups with tumors, i.e. <em>never</em> (no drug), <em>always</em> (pre-drug, i.e. drug treatment before tumor inoculation and throughout the experiment), <em>concurrent</em> (simultaneously tumor inoculation and drug treatment throughout the experiment), and <em>after</em> (post-drug, i.e. drug treatment after tumor inoculation and throughout the experiment). The “psychiatric” conditions of mice were observed from the immobility time with tail suspension and spontaneous motor activity post tumor inoculation. Significant increase of serum interlukin-12 (sIL-12) and the inhibition of tumor growth were found in mirtazapine-treated mice (<em>always</em>, <em>concurrent</em>, and <em>after</em>) as compared with that of <em>never</em>. In addition, interferon-γ level and immunocompetent infiltrating CD4+/CD8+ T cells in the tumors of mirtazapine-treated, tumor-bearing mice were significantly higher as compared with that of <em>never.</em> Tumor necrosis factor-α (TNF-α) expressions, on the contrary, are decreased in the mirtazapine-treated, tumor-bearing mice as compared with that of <em>never</em>. Ex vivo autoradiography with [¹²³I]ADAM, a radiopharmaceutical for serotonin transporter, also confirms the similar results. Notably, better survival rates and intervals were also found in mirtazapine-treated mice. These findings, however, were not observed in the immunodeficient mice. Our results suggest that tumor growth inhibition by mirtazapine in CT26/<em>luc</em> colon carcinoma-bearing mice may be due to the alteration of the tumor microenvironment, which involves the activation of the immune response and the recovery of serotonin level.</p> </div

Body weight loss of the C26/tk-luc colon carcinoma-bearing mice after treatment with various liposomal drugs.

<p>The mice bearing large tumor (<i>n</i> = 9 for each group, A) and those bearing small tumor (<i>n</i> = 6 for each group, B) were injected intravenously with NanoX (•), InNanoX (▽), NanoVNB (▪) or InVNBL (◊) at 0, 7, and 14 days after first injection (arrow; three injections total). The zero time point indicates the initiation of therapy. Data were expressed as mean ± S.E.M.</p

Experimental designs.

<p>(A) Tumor inoculation, mirtazapine treatment, and monitoring of tumor growth and survival. (B) On day 22, mice were assayed for behaviors, then sacrificed for the measurement of lymphocyte subsets and performed with <i>ex vivo</i> autoradiography.</p

Survival fraction of the C26/tk-luc colon carcinoma-bearing mice after treatment with various liposomal drugs.

<p>The mice bearing large tumor (<i>n</i> = 9 for each group, A) and those bearing small tumor (<i>n</i> = 6 for each group, B) were injected intravenously with NanoX (•), InNanoX (▽), NanoVNB (▪) or InVNBL (◊) at 0, 7, and 14 days after first injection (arrow; three injections total). Mice were euthanized when tumor volume greater than 2500 mm³.</p