Tetrahydrofurofuran-type lignans inhibit breast cancer-mediated bone destruction by blocking the vicious cycle between cancer cells, osteoblasts and osteoclasts.

Breast cancer frequently spreads to bone. The interaction between bone metastases and microenvironment, referred as the "vicious cycle", increases both tumor burden and bone destruction. Therefore, inhibition at any point in this "vicious cycle" can reduce malignant osteolytic lesions in patients with advanced breast cancer. In this study, we evaluated whether tetrahydrofurofuran-type lignans derived from Magnoliae Flos, commonly used in traditional Asian medicine to treat inflammatory diseases, could block breast cancer-mediated bone loss. Aschatin, fargesin, lirioresinol B dimethyl ether, and magnolin at noncytotoxic concentrations suppressed mRNA expression and secretion of osteolytic factor PTHrP in MDA-MB-231 metastatic human breast cancer cells. Fargesin inhibited TGF-ß-stimulated cell viability, migration, and invasion and decreased TGF-ß-induced PTHrP production in MDA-MB-231 cells. In addition, these lignans reduced RANKL/OPG ratio in PTHrP-treated hFOB1.19 human osteoblastic cells and inhibited RANKL-mediated osteoclast differentiation in mouse bone marrow macrophages. Aschatin, fargesin, lirioresinol B dimethyl ether, and magnolin substantially reduced bone-resorbing activity of osteoclasts by inhibiting MMP-9 and cathepsin K activities. Furthermore, orally administered fargesin inhibited tumor growth and cancer-mediated bone destruction in mice with MDA-MB-231 cells injected into calvarial tissues. Aschatin, fargesin, lirioresinol B dimethyl ether, and magnolin blocked initiation and progression of the "vicious cycle" between breast cancer metastases and bone microenvironment by inhibiting PTHrP production in breast cancer cells and osteoclastic bone resorption. Therefore, these tetrahydrofurofuran-type lignans have the potential to serve as beneficial agents to prevent and treat cancer-induced bone destruction in breast cancer patients.

[Preparation and identification of recombinant adenoviruses carrying short hairpin RNA targeting parathyroid hormone related protein of goat].

Parathyroid hormone related protein (PTHrP) has important biological functions in calcium metabolism. The aim of this study was to silence the expression of PTHrP by RNA interference and recombinant adenovirus, and to provide a material to investigate the relative functions of PTHrP in goat mammary gland epithelial cell. The Block-iT shRNA interference system was used in this experiment. We designed and synthesized two pairs of complementary single-strand DNA oligonucleotides (shRNA-322/357) targeting two different sites of PTHrP mRNA. Then the oligonucleotides were inserted into shuttle vector pENTR/CMV-GFP/U6. After detection of the interference efficiency by Western blotting, we chose pENTR/CMV-GFP/U6-322 and adenovirus backbone vector pAD/PL-DEST to produce recombinant vector pAD/PL-DEST/CMV-GFP/U6-322. The first generation recombinant adenovirus particles (AD-PTHrP-322) were produced and further amplified by transfecting HEK-293 cells. The titer of the recombinant adenovirus reached 2.0 x 1(9) PFU/mL determined by TCID50 assays. The result of real-time quantitative PCR indicated that mRNA expression levels of gene were reduced 29.2%, 68.1% and 82.6% (P < 0.05), respectively, when goat mammary gland epithelial cells were infected with AD-PTHrP-322 after 24, 48 and 72 h, in which PTHrP. Western blotting also showed that the expression of PTHrP was reduced by infecting the cells with AD-PTHrP-322. AD-PTHrP-322 has been proved with significant interference effect on expression of PTHrP.

Anaplastic thyroid carcinoma with humoral hypercalcemia of malignancy (HHM): an autopsy case report.

An 84-year-old woman was admitted to our hospital for the examination and treatment of painful right thyroid swelling on August 2, 2002. Thyroid ultrasonography showed a mass of about 6 cm in diameter at the right thyroid lobe. Aspiration biopsy cytology (ABC) of her mass showed a thyroid carcinoma. Her neck mass was cold on (123)I scintigraphy and hot on both early- and delayed- phase (201)Tl scintigraphy. Whole body (67)Ga scintigraphy scan showed a strong hot accumulation in the area from the right thyroid lobe to the right lateral lobe. Multiple lung tumors were observed from chest computed tomography (CT) scans. She was diagnosed as having an anaplastic thyroid carcinoma with metastatic lung tumors. As her thyroid carcinoma was inoperable, percutaneous injection therapy of lipiodol and ethanol (lip-PEIT) against the primary thyroid carcinoma was performed twice a week. However, the thyroid carcinoma gradually enlarged and oppressed her trachea. Two months after the initiation of lip-PEIT, parathyroid hormone-related protein (PTHrP)-dependent hypercalcemia was diagnosed because serum levels of calcium, phosphate and intact-PTHrP were 2.72 mmol/l (10.9 mg/dl), 0.71 mmol/l (2.2 mg/dl), 3.2 pmol/l, respectively. The hypercalcemia was reduced by the use of pamidronate. After one week she died of an airway obstruction caused by the developing thyroid carcinoma. Carcinoma cells with a mixed papillary and squamoid pattern were positively stained immunohistochemically by anti-PTHrP(1-34) antisera. Herein, we report a rare autopsy case of a PTHrP-producing thyroid carcinoma.