The inhibitory effects of capillarisin on cell proliferation and invasion of prostate carcinoma cells

Objectives

Capillarisin (Cap), an active component of Artemisia capillaris root extracts, is characterized by its anti‐inflammatory, anti‐oxidant and anti‐cancer properties. Nevertheless, the functions of Cap in prostate cancer have not been fully explored. We evaluated the potential actions of Cap on the cell proliferation, migration and invasion of prostate carcinoma cells.

Materials and methods

Cell proliferation and cell cycle distribution were measured by water‐soluble tetrazolium‐1 and flow cytometry assays. The expression of cyclins, p21, p27, survivin, matrix metallopeptidase (MMP2 and MMP9) were assessed by immunoblotting assays. Effects of Cap on invasion and migration were determined by wound closure and matrigel transmigration assays. The constitutive and interlukin‐6 (IL‐6)‐inducible STAT3 activation of prostate carcinoma cells were determined by immunoblotting and reporter assays.

Results

Capillarisin inhibited androgen‐independent DU145 and androgen‐dependent LNCaP cell growth through the induction of cell cycle arrest at the G0/G1 phase by upregulating p21 and p27 while downregulating expression of cyclin D1, cyclin A and cyclin B. Cap decreased protein expression of survivin, MMP‐2, and MMP‐9 and therefore blocked the migration and invasion of DU145 cells. Cap suppressed constitutive and IL‐6‐inducible STAT3 activation in DU145 and LNCaP cells.

Conclusions

Our data indicate that Cap blocked cell growth by modulation of p21, p27 and cyclins. The inhibitory effects of Cap on survivin, MMP‐2, MMP‐9 and STAT3 activation may account for the suppression of invasion in prostate carcinoma cells. Our data suggest that Cap might be a therapeutic agent in treating advanced prostate cancer with constitutive STAT3 or IL‐6‐inducible STAT3 activation.

     

Dysanaptic growth of conducting airways after pneumonectomy assessed by CT scan.

In immature dogs after pneumonectomy (PNX), pulmonary viscous resistance is persistently elevated predominantly as a result of a high airway resistance (Raw). We examined the anatomical basis for this observation by using computerized tomography scans obtained from foxhounds 4-10 mo after right PNX. Airways of the left lower lobe were followed from generations z = 0 (trachea) to z = 12. By 4 mo post-PNX, airway length increased significantly relative to sham-operated dogs, but airway cross-sectional area (CSA) did not. By 10 mo post-PNX, average airway CSA was 24% above that in controls. Theoretically, the increased airway length and CSA should reduce lobar Raw by 50%. However, post-PNX airway dilatation did not normalize total CSA, and estimated resistance due to turbulence and convective acceleration increased threefold; i.e., the 50% reduction in lobar Raw would be offset by the loss of four of seven lobes. Thus the expected reduction in work of breathing in the whole animal is only ~30%, consistent with previously measured work of breathing in pneumonectomized dogs. We conclude that airway structure adapts slowly and incompletely, resulting in limited functional compensation.     

Inhibition of lipid synthesis and glucose-6-phosphate dehydrogenase in rat skin by dehydroepiandrosterone

Lipid synthesis from acetate-1-(14)C by rat skin was inhibited 44-56% by 2.5 x 10(-4) m dehydroepiandrosterone (DHA) in vitro with or without addition of glucose in the incubation medium. This inhibition affected all the lipid fractions examined (hydrocarbons, sterols, sterol esters, tri-, di- and monoglycerides, fatty acids, and polar lipids) and could be reversed by NADPH. DHA also inhibited lipid synthesis from glucose-U-(14)C and the formation of (14)CO(2) from glucose-1-(14)C, indicating interference with pentose cycle activity. Experiments with the 105,000 g supernatant fluid of rat skin homogenates demonstrated considerable activities of malic enzyme (ME) (12.6 nmoles of NADPH generated per min per mg of protein), of glucose-6-phosphate dehydrogenase (G6PD), and of 6-phosphogluconate dehydrogenase (6PGD) (17.5 nmoles of NADPH generated per min per mg of protein). G6PD was inhibited 98% by 2.5 x 10(-4) m dehydroepiandrosterone, while 6PGD and ME were not affected. It can be estimated from these data that the pentose cycle may contribute 41-57% of the NADPH needed for lipid synthesis in rat skin; the remainder of the necessary NADPH is presumably supplied by malic enzyme.     

Purification and characterization of genetic variants of 6-phosphogluconate dehydrogenase

This paper describes the physicochemical characteristics in partially purified enzyme on subjects with the Pd A, Pd AB, and Pd B variants of 6-phosphogluconic dehydrogenase (6PGD). For these studies, whole blood was purified about 225-fold using ion exchange chromatography on DEAE cellulose column and fractionation with ammonium sulfate. 6PGD emerges as a single peak between 0.01 m and 0.1 m phosphate buffer on the column and is precipitated in the 55–80% fraction of ammonium sulfate. This purified enzyme can be stored frozen for several months without appreciable loss of activity and contains no detectable activity of glucose 6-phosphate dehydrogenase and glutathione reductase. The three variants of partially purified 6PGD varied from each other in two respects. The transitional temperature is 47.8 C for Pd A, 45.4 C for Pd AB, and 41.1 C for Pd B. The K _m for 6PGA is 30 m for Pd A, 21 m for Pd AB, and 15 mfor Pd B. These observations add further strength to the concept that the polymorphism in 6PGD represents alterations in either the configuration or structure of the protein molecule itself.Supported by grants from the Chicago Community Trust and the U.S. Public Health Service (Tl-AM-5186).     

Red cell distribution and the recruitment of pulmonary diffusing capacity.

The distribution of red blood cells in alveolar capillaries is typically nonuniform, as shown by intravital microscopy and in alveolar tissue fixed in situ. To determine the effects of red cell distribution on pulmonary diffusive gas transport, we computed the uptake of CO across a two-dimensional geometric capillary model containing a variable number of red blood cells. Red blood cells are spaced uniformly, randomly, or clustered without overlap within the capillary. Total CO diffusing capacity (DLCO) and membrane diffusing capacity (DmCO) are calculated by a finite-element method. Results show that distribution of red blood cells at a fixed hematocrit greatly affects capillary CO uptake. At any given average capillary red cell density, the uniform distribution of red blood cells yields the highest DmCO and DLCO, whereas the clustered distribution yields the lowest values. Random nonuniform distribution of red blood cells within a single capillary segment reduces diffusive CO uptake by up to 30%. Nonuniform distribution of red blood cells among separate capillary segments can reduce diffusive CO uptake by >50%. This analysis demonstrates that pulmonary microvascular recruitment for gas exchange does not depend solely on the number of patent capillaries or the hematocrit; simple redistribution of red blood cells within capillaries can potentially account for 50% of the observed physiological recruitment of DLCO from rest to exercise.     

Cytomegalovirus (CMV) DNA Load Is an Independent Predictor of CMV Disease and Survival in Advanced AIDS

The impact of cytomegalovirus (CMV) on human immunodeficiency virus type 1 (HIV-1) disease progression has been controversial. In this study, we sought to determine if CMV viral load is independent of HIV-1 viral load in predicting CMV disease and survival. Our findings indicate that in patients with advanced AIDS, CMV DNA load is an independent marker of CMV disease and survival and is more predictive than HIV-1 RNA load. Moreover, patients who respond to preemptive therapy with oral ganciclovir, with resulting undetectable levels of CMV DNA, in their plasma, have a significantly lower risk of developing CMV disease and higher rates of survival, despite stable or increasing HIV-1 RNA loads. These data provide support for CMV as an independent risk factor for mortality in persons with advanced AIDS and further suggest that effective preemptive therapy for CMV can improve patient survival rates.