Lung Collagen Hydroxyproline Research Articles

AS1411 aptamer tagged PEGylated liposomes as a smart nanocarrier for tumor-specific delivery of Withaferin A for mitigating pulmonary metastasis

Metastasis is the most challenging health problem contributing to about 90 % of cancer-related deaths worldwide. Metastatic tumors are highly aggressive and resistant to the most available therapeutic options. Hence, innovative therapeutic approaches are required to target metastatic tumors selectively. In this study, we prepared AS1411 functionalized Withaferin A loaded PEGylated nanoliposomes (ALW) and investigated its therapeutic effect in B16F10 induced in pulmonary metastasis mice models. The prepared formulations' size and morphological properties were evaluated using dynamic light scattering system and Transmission electron microscope. ALW had spherical-shaped nanosized particles with a size of 118 nm and an encapsulation efficacy of 82.5 %. TEM analysis data indicated that ALW has excellent dispersibility and uniform spherical nano-size particles. ALW inhibited cell viability, and induced cell apoptosis of B16F10. In vivo, the pulmonary metastasis study in C57BL/6 mice revealed that the ALW significantly (p < 0.01) improved the encapsulated WA anti-metastatic activity and survival rate compared to WA or LW treated groups. ALW significantly (p < 0.01) downregulated the levels of IL-6, TNF-α, and IL-1β and significantly reduced the lung collagen hydroxyproline, hexosamine, and uronic acid content in metastatic tumor bearing animals compared to WA or LW. Gene expression levels of MMPs and NF-κB were downregulated in ALW treated metastatic pulmonary tumor-bearing mice. These findings demonstrate that the AS1411 functionalized Withaferin A loaded PEGylated nanoliposomes could be a promising nanoliposomal formulation for targeting metastatic tumors.

Targeted inhibition of tumor survival, metastasis and angiogenesis by Acacia ferruginea mediated regulation of VEGF, inflammatory mediators, cytokine profile and inhibition of transcription factor activation

Acacia ferruginea extract (AFE) was studied for anti-metastasis/-angiogenesis activity against B16F-10 melanoma cells in C57BL/6 mice. In vitro cytotoxicity of AFE was first screened using MTT assay and it was shown to inhibit B16F-10 cells with IC50 value of 52.94 μg/ml. Anti-metastatic activity of AFE in vivo revealed administration of AFE (10 mg/kg.b.wt) in three different regimens has shown reduced metastatic colony formation in lungs and prolonged survival in metastatic tumor-bearing hosts. Biochemical analysis shown that treatment with AFE significantly reduced the lung collagen hydroxyproline, hexosamine, uronic acid, sialic acid and gamma-glutamyl transpeptidase content. Administration of AFE significantly inhibited the iNOS and COX-2 level, diminished the infiltration of neoplastic cells and subsequently reduced the number of p53 and Bcl-2 positive immunoreactive cells as evidenced by histological and immunohistochemistry analysis. In addition, we found that, AFE significantly decreased the level of pro-inflammatory cytokines interleukin-1β, IL-6, TNF-α and suppressed the nuclear factor kappa B (NF-κB) activation. Furthermore, angiogenesis studies shown significant reduction in number of tumor directed capillaries, regulating cytokines and vascular endothelial growth factor (VEGF). Our present results suggests that AFE could be a promising candidate for developing anti-cancer agents targeting metastasis which helps further to combat melanoma with effective treatment strategy.

Inhibition of pulmonary metastasis by Emilia sonchifolia (L.) DC: An in vivo experimental study

BackgroundEmilia sonchifolia (L.) DC is a widely distributed medicinal herb used mainly in the indigenous Ayurvedic system of medicine in India. This plant is one among the ten sacred plants of Kerala state in India, collectively known as Dasapushpam. PurposeTo assess the therapeutic efficacy of this well-known medicinal plant in a catastrophic complication like metastatic cancer progression. This study further aimed to scientifically validate the traditional medicinal use of this sacred plant. Study designHighly metastatic B16F10 melanoma will spontaneously metastasize in C57BL/6 mice and is accepted as a useful murine model for the study on metastasis. Three different experimental modalities of prophylactic, simultaneous and after tumour development were used for data accumulation and analysis. MethodsWhole plant genuine extract of E. sonchifolia (25 mg/kg bodyweight) was administered intraperitoneally to C57BL/6 mice. Animals were sacrificed on 21st day after tumour induction and the lung tumour nodules were counted. Various lung and serum biochemical parameters along with major cytokine levels were recorded. Survival rate was monitored. Histopathology of the lung tissue and expression studies of the major genes involved in metastasis was also carried out. ResultsE. sonchifolia significantly inhibited pulmonary tumour formation and increased the life span of animals. Lung collagen hydroxyproline, uronic acid, hexosamine, serum sialic acid, γ-glutamyl transpeptidase, vascular endothelial growth factor (VEGF), granulocyte monocyte colony-stimulating factor and other cytokine levels were significantly lowered in the treated group of animals. Histopathological analysis was also correlated with these findings. E. sonchifolia down regulated the expression of matrix metalloproteinases; extracellular signal-regulated kinases and VEGF at the same time up regulated the expression of tissue inhibitor of matrix metalloproteinases. ConclusionPrevious studies on E. sonchifolia proved its significant biological properties including anti-tumour, anti-inflammatory and antioxidant activities. Present report is so far the first study to demonstrate the anti-metastatic potential of this medicinal herb justifying its conventional use in the traditional medicine.

Protective Effect of Solanum muricatum on Tumor Metastasis by Regulating Inflammatory Mediators and Nuclear Factor-Kappa B Subunits

Metastasis is one of the hallmarks of malignant neoplasm or cancer, which is the leading cause of death in many cancer patients. A major challenge in cancer treatment is to find better ways to specifically target tumor metastases. In this study, the anti-metastatic potential of the methanol extract of Solanum muricatum (S. muricatum) was evaluated using B16F-10 melanoma cell-induced lung metastasis in C57BL/6 mice. Treatment with S. muricatum significantly inhibited the lung tumor nodule formation and reduced the lung collagen hydroxyproline, hexosamine, and uronic acid levels (p<0.01). Similarly, serum sialic acid and γ-glutamyl transpeptidase levels were also significantly inhibited after S. muricatum treatment. The levels of proinflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, granulocyte monocyte colony-stimulating factor (GM-CSF), and IL-2 in serum were significantly regulated after treatment with S. muricatum. The serum nitric oxide level was also decreased significantly and was accompanied by a decrease in inducible nitric oxide synthase (iNOS) and cyclo- oxygenase (COX)-2 expressions after S. muricatum treatment. The present study reveals that S. muricatum treatment was able to alter the proinflammatory cytokine production as well as inhibit the activation and nuclear translocation of nuclear factor-κB (p65 and p50) subunits.

Decalepis hamiltonii Inhibits Tumor Progression and Metastasis by Regulating the Inflammatory Mediators and Nuclear Factor κB Subunits

Metastasis is an extremely complex process that is a major problem in the management of cancer. In the present study, we had evaluated the antimetastatic activity of DECALEPIS HAMILTONI: using B16F-10 melanoma-induced experimental lung metastasis in a C57BL/6 mice model. treatment significantly ( : < .01) inhibited lung tumor nodule formation and reduced the lung collagen hydroxyproline, hexosamine, and uronic acid levels. Similarly serum sialic acid and γ-glutamyl transpeptidase levels were also significantly inhibited after treatment. The levels of proinflammatory cytokines such as tumor necrosis factor α, interleukin (IL)-1β, IL-6, granulocyte monocyte colony-stimulating factor, and IL-2 in the serum of these animals were significantly altered after treatment. The serum NO level was also found to be significantly decreased after treatment. This decreased NO level after treatment was also accompanied by decreased inducible NO synthase and cyclooxygenase-2 expression. The study reveals that treatment could alter proinflammatory cytokine production and could inhibit the activation and nuclear translocation of p65 and p50 subunits of nuclear factor κB in B16F-10 cells.

Inhibition of Metastatic Lung Cancer in C57BL/6 Mice by Marine Mangrove Rhizophora apiculata

Metastasis is one of the hallmarks of malignant neoplasms and is the leading cause of death in many cancer patients. A major challenge in cancer treatment is to find better ways to specifically target tumor metastasis. In this study, the anti-metastatic potential of the methanolic extract of Rhizophora apiculata (R.apiculata) was evaluated using the B16F-10 melanoma induced lung metastasis model in C57BL/6 mice. Metastasis was induced in C57BL/6 mice by injecting highly metastatic B16F-10 melanoma cells through the lateral tail vein. Simultaneous treatment with R.apiculata extract (10 mg/kg b.wt (intraperitoneal) significantly (p<0.01) inhibited pulmonary tumor nodule formation (41.1 %) and also increased the life span (survival rate) 107.3 % of metastatic tumor bearing animals. The administration of R.apiculata extract significantly (p<0.01) reduced biochemical parameters such as lung collagen hydroxyproline, hexosamine, uronic acid content, serum nitric oxide (NO), γ-glutamyl transpeptidase (GGT) and sialic acid levels when compared to metastasis controls. These results correlated with lung histopathology analysis of R.apiculata extract treated mice showing reduction in lung metastasis and tumor masses. Taken together, our findings support that R.apiculata extract could be used as a potential anti-metastasis agent against lung cancer.

Inhibition of B16F-10 Melanoma–Induced Lung Metastasis in C57BL/6 Mice by Aerva lanata via Induction of Apoptosis

In this study, the antimetastatic potential of the ethanolic extract of Aerva lanata was evaluated using the B16F-10 melanoma-induced lung metastasis model. Metastasis was induced in C57BL/6 mice by injecting highly metastatic B16F-10 melanoma cells through the lateral tail vein. Simultaneous treatment with A lanata inhibited tumor nodule formation in the lungs (70.53%), and there was a 65.3% increase in the survival rate of metastatic tumor-bearing animals. These results correlated with biochemical parameters such as lung collagen hydroxyproline, hexosamine, and uronic acid contents; serum sialic acid and γ-glutamyl transpeptidase levels; and histopathological analysis. In vitro studies using B16F-10 cells showed that A lanata inhibited migration of tumor cells, cell invasion through type-I collagen-coated polycarbonate filter and activation of matrix metalloproteinases. Treatment with A lanata induced apoptotic response, characterized by apoptotic morphology, a typical ladder of DNA fragmentation, and detection of 3' hydroxyl ends in DNA by TUNEL assay. There was an increase in the percentage of cells in the sub-G0/G1 phase indicating cell cycle arrest. A lanata treatment resulted in downregulation of bcl-2 and cyclin-D1 expression and upregulation of p53, bax, caspase-9, caspase-3, p21, and p27 gene expression in B16F-10 cells. Proinflammatory cytokine production and gene expression were also found to be downregulated in A lanata-treated cells.

Thujone inhibits lung metastasis induced by B16F-10 melanoma cells in C57BL/6 mice

The antimetastatic potential of thujone, a naturally occurring monoterpene, was evaluated. Metastasis was induced in C57BL/6 mice by injecting highly metastatic B16F-10 melanoma cells through the lateral tail vein. Administration of thujone (1 mg·(kg body weight)(-1)), prophylactically and simultaneously with tumor induction, inhibited tumor nodule formation in the lungs by 59.45% and 57.54%, respectively, with an increase in the survival rate (33.67% and 32.16%) of the metastatic tumor bearing animals. These results correlated with biochemical parameters such as lung collagen hydroxyproline, hexosamine and uronic acid contents, serum sialic acid and γ-glutamyl transpeptidase levels, and histopathological analysis. Treatment with thujone downregulated the production of proinflammatory cytokines such as tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and granulocyte-monocyte colony-stimulating factor. Thujone administration downregulated the expression of matrix metalloproteinase (MMP)-2, MMP-9, extracellular signal-regulated kinase (ERK)-1, ERK-2, and vascular endothelial growth factor (VEGF) and also upregulated the expression of nm-23, tissue inhibitor of metalloproteinase (TIMP)-1, and TIMP-2 in the lung tissue of metastasis-induced animals. Treatment with thujone inhibited the activity of MMP-2 and MMP-9 in gelatin zymographic analysis. Thujone treatment significantly inhibited the invasion of B16F-10 melanoma cells across the collagen matrix in a Boyden chamber. Thujone also inhibited the adhesion of tumor cells to collagen-coated microtire plate wells and the migration of B16F-10 melanoma cells across a polycarbonate filter in vitro. These results indicate that Thujone can inhibit the lung metastasis of B16F-10 cells through inhibition of tumor cell proliferation, adhesion, and invasion, as well as by regulating expression of MMPs, VEGF, ERK-1, ERK-2, TIMPs, nm23, and levels of proinflammatory cytokines and IL-2 in metastatic animals.

Vernonia cinerea Less. Inhibits Tumor Cell Invasion and Pulmonary Metastasis in C57BL/6 Mice

The effect of Vernonia cinerea Less. extract on the inhibition of lung metastasis induced by B16F-10 melanoma cells was studied in C57BL/6 mice. V cinerea extract significantly (P < .001) inhibited lung tumor formation (78.8%) and significantly increased the life span (72.5%). Moreover, lung collagen hydroxyproline, uronic acid, and hexosamine and also serum sialic acid, γ-glutamyltransferase (GGT), and vascular endothelial growth factor (VEGF) levels were found to be significantly (P < .001) lower in treated animals compared with untreated controls. Histopathological analysis of the lung tissues also correlated with these findings. V cinerea treatment significantly inhibited the invasion of B16F-10 melanoma cells across the collagen matrix of the Boyden chamber. V cinerea also inhibited the migration of B16F-10 melanoma cells across a polycarbonate filter in vitro. It downregulated the production and expression of proinflammatory cytokines such as TNF (tumor necrosis factor)-α, IL (interleukin)-1β, IL-6, and GM-CSF (granulocyte monocyte colony-stimulating factor). V cinerea extract administration could suppress or downregulate the expression of matrix metalloproteinase (MMP)-2, MMP-9, lysyl oxidase, prolyl hydroxylase, K-ras, extracellular signal-regulated kinase (ERK)-1, ERK-2, and VEGF and also upregulate the expression of nm-23, tissue inhibitor of metalloproteinase (TIMP-1), and TIMP-2 in the lung tissue of metastasis-induced animals. It also inhibited the protein expression of MMP-2 and MMP-9 in gelatin zymographic analysis of B16F-10 cells. These results indicate that V cinerea could inhibit the metastatic progression of B16F-10 melanoma cells in C57BL/6 mice by regulating MMPs, VEGF, prolyl hydroxylase, lysyl oxidase, ERK-1, ERK-2, TIMPs, nm23, and proinflammatory cytokine gene expression in metastatic lung tissue.

Effect of Amentoflavone on the Inhibition of Pulmonary Metastasis Induced by B16F-10 Melanoma Cells in C57BL/6 Mice

This study was an investigation of the antimetastatic activity of amentoflavone using B16F-10 melanoma-induced experimental lung metastasis in C57BL/6 mice. Amentoflavone treatment significantly reduced tumor nodule formation accompanied by reduced lung collagen hydroxyproline, hexosamine, and uronic acid levels. Serum sialic acid and gammaglutamyl transpeptidase levels were also significantly inhibited after amentoflavone treatment. Amentoflavone treatment up-regulated the lung tissue inhibitor of metalloprotease-1 and tissue inhibitor of metalloprotease-2 expression. The cytokine profile and growth factors such as interleukin-1beta , interleukin-6, tumor necrosis factor-alpha, granulocyte monocyte- colony stimulating factor, vascular endothelial growth factor, interleukin-2, and tissue inhibitor of metalloprotease-1 in the serum of these animals were markedly altered after amentoflavone treatment. This altered level of cytokines after amentoflavone treatment was also accompanied by enhanced natural killer cell antibody-dependent cellular cytotoxicity. The study reveals that amentoflavone treatment could alter proinflammatory cytokine production and could inhibit the activation and nuclear translocation of p65, p50, c-Rel subunits of nuclear factor-kappaB, and other transcription factors such as c-fos, activated transcription factor-2, and cyclic adenosine monophosphate response element-binding protein in B16F-10 melanoma cells.

A Preliminary Study on Antimetastatic Activity of Thuja Occidentalis L. in Mice Model

The effect of Thuja occidentalis extract on the inhibition of lung metastasis induced by B16F-10 melanoma cells was studied in C57BL/6 mice. The extract was administered by three different modalities. A remarkable reduction in tumor-nodule formation was shown by simultaneous (74.4%) and prophylactic (71.5%) mode of administration. The effect was comparatively low in drug administration after tumor development (60.2%). Increased lung collagen hydroxyproline (21.13 µg/mg protein) in the metastasized lungs of control animals compared with normal animals (0.98 µg/mg protein) was significantly reduced in Thuja-treated animals. The elevated level of uronic acid (349.5 µg/100 mg tissue) and hexosamine contents in metastatic control animals was significantly reduced in the animals treated with alcoholic extract of Thuja. Similarly the elevated levels of serum sialic acid and serum gamma glutamyl transpeptidase activity in the untreated control animals was significantly reduced in the animals treated with the extract of Thuja. The lifespan of the Thuja treated animals also was seen to be significantly increased.

Inhibitory effect of Boerhaavia diffusa on experimental metastasis by B16F10 melanoma in C57BL/6 mice

Administration of the aqueous methanol (3:7) extract of B.diffusa was found to be effective in reducing the metastases formation by B16F10 melanoma cells. Prophylactic administration of the extract (0.5mg/dose) inhibited the metastases formation by about 95% as compared to untreated control animals. There was 87% of inhibition in the lung metastases formation in syngenic C57BL/6 mice, when the extract was administered simultaneously with tumour challenge. Biochemical parameters such as lung collagen hydroxyproline, hexosamines and uronic acid levels were also reduced significantly (P < 0.001) in the treated animals. Levels of serum sialic acids and γ-glutamyltranspeptidase that are markers of neoplastic proliferation were also reduced in the tumour plus extract treated animals. More over treatment with the extract enhanced the survival of the animals more than double that of untreated control animals. When a non-toxic concentration of the extract was treated directly to the B16F10 cells in vitro, it inhibited the cell proliferation as estimated by the 3H - thymidine uptake assay. From the Zymogram analysis using culture supernatant from the extract treated cells it became evident that the components of the extract inhibited the expression or activity of gelatinases A and B (MMP-2 and MMP-9). Since the MMPs are intimately associated with cell invasion and angiogenesis, inhibition of these functions along with the anti-proliferative activity (cytostatic) may be contributing to the antimetastatic property shown by B. diffusa.

Effect of Withania somnifera on B16F‐10 melanoma induced metastasis in mice

Withania somnifera, a plant with known immunopotentiating activity and its bioactive fraction-Withanolide D were studied for their anti-metastatic activity using B16F-10 melanoma cells in C57BL/6 mice. Simultaneous administration of Withania extract (122 +/- 10 tumour nodules) and Withanolide (126 +/- 9 lung tumour nodules) could significantly (p < 0.001) inhibit the metastatic colony formation of the melanoma in lungs. 72.58% by extract and 69.84% by Withanolide treated, as compared to the untreated control animals also increased the survival days. Lung collagen hydroxyproline content was highly elevated in the control animals (23.5 +/- 0.9 micro g/mg protein), which was reduced by the simultaneous administration of both the extract (16.3 +/- 2.0 micro g/mg protein) and Withanolide (15.3 +/- 1.8 micro g/mg protein). The level of lung hexosamines (4.85 +/- 0.20 mg/100 mg tissue) and uronic acids (330.1 +/- 23.7 micro g/100 mg tissue) content was also elevated in the control animals. The elevated level of hexosamine was significantly reduced by the treatment with extract (1.92 +/- 0.05) and Withanolide (1.85 +/- 0.05). Similarly, the uronic acid content was also been reduced by the simultaneous administration of both Withania extract (194.2 +/- 17.4) and Withanolide (183.2 +/- 8.8). The control animals had 35.3 +/- 3.8 U/L gamma-glutamyl transpeptidase (gamma-GT), which was reduced by 50% by the treatment of extract and Withanolide to 17.5 +/- 4.0 U/L and 16.3 +/- 4.4 U/L respectively. There was a significant reduction in the levels of sialic acid in the serum of Withania extract (60.7 +/- 7.7) and Withanolide (67.16 +/- 5.8) treated animals compared to the higher level (102.2 +/- 8.7) in the control animals. Histopathological analysis of the lung tissues also correlated with these findings. Prophylactic administrations of both extract as well as Withanolide were ineffective in inhibiting the metastasis of B16F-10 melanoma cells.

Inhibitory effect of a polysaccharide from Tinospora cordifolia on experimental metastasis

Administration of the polysaccharide fraction from Tinospora cordifolia was found to be very effective in reducing the metastatic potential of B16F-10 melanoma cells. There was a 72% inhibition in the metastases formation in the lungs of syngeneic C57BL/6 mice, when the drug was administered simultaneously with tumour challenge. Biochemical parameters such as lung collagen hydroxyproline, hexosamines and uronic acids that are markers of neoplastic development were reduced significantly ( P<0.001) in the treated animals compared with the untreated control animals. The treatment could also reduce serum γ-glutamyltranspeptidase (γ-GT) and sialic acid levels as compared to the control animals.

Effect of naturally occurring allyl and phenyl isothiocyanates in the inhibition of experimental pulmonary metastasis induced by B16F-10 melanoma cells

The effect of naturally occurring allyl and phenyl isothiocyanates in the inhibition of B16F-10 melanoma cells-induced metastasis has been studied in C57BL/6 mice. The two compounds were administered by three different modalities. Simultaneous administration of the compounds with tumour induction was found to produce significant reduction in the lung tumour nodule formation; of the two compounds studied, allyl isothiocyanate have shown to be more effective and it produced 93% inhibition of tumour nodules while phenyl isothiocyanate has an inhibition of 77.3%. Prophylactic administration also produces considerable effect but no effect was observed by their administration after the tumour development. Lung collagen hydroxyproline, lung uronic acid, lung hexosamine content, serum sialic acid and serum GGT levels were also estimated and the enhanced levels of these parameters in the control animals were found to be reduced by the simultaneous administration of these isothiocyanates; these data were also correlated with the inhibition of lung tumour nodules produced and the increase in life span of the animals.

Effect of beta-carotene on the inhibition of lung metastasis in mice.

Effect of beta-carotene on the inhibition of lung metastasis induced by B16F-10 melanoma cells was studied in C57BL/6 mice. Simultaneous administration of the compound after tumor induction produced a significant reduction (71.36%) in tumor nodule formation. Increased lung collagen hydroxyproline (22.37 microg/mg protein) in the metastasized lungs of control animals compared to the normal animals (0.95 microg/mg protein) was significantly reduced (4.19 microg/mg protein) in the beta-carotene treated animals. High amount of uronic acid (355.83 microg/100mg tissue ) in the metastasized control animals was significantly reduced (87.87 microg/100 mg tissue) in the animals treated with beta-carotene. Lung hexosamine content also was inhibited significantly in the beta-carotene treated animals (1.58 mg/100 mg lyophilized tissue) compared to the untreated control animals (4.2 mg/100 mg lyophilized tissue). The elevated levels of serum sialic acid and serum gamma glutamyl transpeptidase activity in the untreated control animals was significantly reduced in the animals treated with beta-carotene. Beta carotene treated animals were survived up to 69 days. Histopathology of the lung tissue also correlated with the above parameters and life span of the drug treated animals. Our results reveal the antimetastatic activity of beta-carotene which are abundantly present in green plants, vegetables and fruits.

Effect of piperine on the inhibition of lung metastasis induced B16F-10 melanoma cells in mice.

The effect of piperine on the inhibition of lung metastasis induced by B16F-10 melanoma cells was studied in C57BL/6 mice. Simultaneous administration of the compound with tumor induction produced a significant reduction (95.2%) in tumor nodule formation. Increased lung collagen hydroxyproline (22.37 microg/mg protein) in the metastasized lungs of the control animals compared to normal animals (0.95 microg/mg protein) was significantly reduced (2.59 microg/mg protein) in the piperine-treated animals. The high amount of uronic acid (355.83 microg/100 mg tissue) in the metastasized control animals was significantly reduced (65 microg/100 mg tissue) in the animals treated with piperine. Lung hexosamine content was also significantly reduced in the piperine-treated animals (0.98 mg/100 mg lyophilized tissue) compared to the untreated tumor-bearing animals (4.2 mg/100 mg lyophilized tissue). The elevated levels of serum sialic acid and serum gamma glutamyl transpeptidase activity in the untreated control animals was significantly reduced in the animals treated with piperine. The piperine-treated animals even survived the experiment (90 days). Histopathology of the lung tissue also correlated with the lifespan of the drug-treated animals. Our results demonstrate the antimetastatic activity of piperine, an alkaloid present in plants such as Piper nigrum and Piper longum.

Anti-metastatic activity of curcumin and catechin

The inhibitory effects of curcumin and catechin on lung metastasis induced by B16F-10 melanoma cells were studied in female C57BL/6 mice. Curcumin and catechin significantly ( P<0.001) inhibited lung tumour formation (89.3% and 82.2%, respectively) and significantly increased the life span (143.9% and 80.8%, respectively). Moreover, lung collagen hydroxyproline and serum sialic acid levels were found to be significantly ( P<0.001) lower in treated animals compared to the untreated controls. Curcumin and catechin treatment (10 μg/ml) significantly inhibited the invasion of B16F-10 melanoma cells across the collagen matrix of the Boyden chamber. Gelatin zymographic analysis of the trypsin-activated B16F-10 melanoma cells sonicate revealed that curcumin- and catechin-treated zymograms did not show any metalloproteinase activity. Curcumin and catechin treatment did not inhibit the motility of B16F-10 melanoma cells across a polycarbonate filter in vitro. These findings suggest that curcumin and catechin inhibit the invasion of B16F-10 melanoma cells by inhibition of metalloproteinases, thereby inhibiting lung metastasis.

Effect of Diallyl Sulphide, Diallyl Disulphide, and Allyl Methyl Sulphide on the Inhibition of Lung Metastasis of B16F-10 Melanoma Cells in Mice.

Effect of naturally occurring sulphur-containing compounds such as diallyl sulphide (DAS), diallyl disulphide (DADS), and allyl methyl sulphide (AMS) on the inhibition of lung metastasis of B16F-10 melanoma cells was studied in C57BL/6 mice. Simultaneous administration of the compounds after tumor induction produced a significant reduction in tumor nodule formation (DAS 55%, DADS 90%, and AMS 54%). The effect was lower when the compounds were administered after tumor development, whereas prophylactic administration of the compounds did not have any effect on lung tumor nodule formation. Increased lung collagen hydroxyproline, serum sialic acid, and gamma glutamyl transpeptidase activity in the metastasized lungs of control animals was significantly reduced in the animals treated with the sulphur-containing compounds. Pathology of the lung tissue also correlated with the above findings. Our results are indicative of the antimetastatic activity of these sulphur-containing compounds.

Inhibition of lung metastasis by adoptive immunotherapy using iscador

Adoptive immunotherapy using spleen cells activated with Iscador was found to inhibit tumour growth significantly (P<0.001). Metastatic B16F10 tumour bearing animals treated with a single dose of spleen cells activated with Iscador (in vitro) showed 100% inhibition of tumour nodule formation on 21st day. Single injection of splenocytes isolated from mice treated with Iscador inhibited the tumour nodule formation by 93.8%. Animals treated with in vivo and in vitro activated spleen cells along with Iscador had an increase in life span of 119% and 81% respectively. Treatment of animals with low dose of Iscador after adoptive immunotherapy further augmented the life span. Animals which underwent adoptive immunotherapy showed significantly reduced lung collagen hydroxyproline (9.8μg/mg protein) and serum sialicacid (24 6μg/ml serum) levels compared to control tumour bearing animals with increased levels of lung hydroxyproline (26.95μg/mg protein) and serum sialic acid levels (151.3μg/ml serum) Serum γ -glutamyl transpeptidase levels were found to be significantly reduced in the group of animals treated with Iscador and Iscador activated splenocytes (16.6 ± 2.3 nmol P-nitroaniline released/ml serum) Group of animals treated with Iscador activated splenocytes alone also showed significantly reduced serum γ -glutamyl transpeptidase levels (17.3 ± 10 nmol P-nitroaniline released/ml serum) compared to control tumour bearing animals (91 • 12 nmol P-nitroaniline released /ml serum)