Screening antitumor bioactive fraction from Sauromatum giganteum (Engl.) Cusimano & Hett and sensitive cell lines with the serum pharmacology method and identification by UPLC-TOF-MS.

Sauromatum giganteum (Engl.) Cusimano & Hett Tuber are used in Chinese folklore medicine for treatment of neoplasms. However, the claim has not been scientifically validated. The aim of the study is to screen the antitumor bioactive fraction of Sauromatum giganteum (Engl.) Cusimano & Hett Tuber and sensitive tumor cell lines using a cytotoxicity assay in vitro and tumor transplantation method in vivo, to support its use in folk medicine. The petroleum ether fraction, chloroform fraction, ethyl acetate fraction, n-butanol fraction and water fraction were successively extracted by turn by the maceration under reflux assay. Screening of antitumor bioactive fraction and sensitive cell lines were measured by MTT assay and the serum pharmacology method, and in vivo the antitumor activities of the active fraction was evaluated by using S180 or H22 tumor-bearing mice model and Kunming mice. The active constituents of ethyl acetate fraction of Sauromatum giganteum (Engl.) Cusimano & Hett were characterized by UPLC-TOF-MS. Compared with control groups, mice serum containing ethyl acetate fraction had a inhibition effect on SMMC-7721 cell, SGC-7901 cell, MCF-7 cell, HeLa cell, A549 cell, HT-29, and MDA-MB-231, respectively, but mice serum containing other four fractions had no different with that of control group. The inhibition capabilities of mice serum containing ethyl acetate fraction on the seven cell lines in descending order is SGC-7901 > SMMC-7721 > MCF-7 > HT-29 > A549 > HeLa > MDA-MB-231. In vivo the inhibition rate of 106, 318, 954 mg/kg·d ethyl acetate fraction dry extract to sarcoma S180 is 15.22%, 26.15% and 40.24%, respectively, and life prolonging rate to hepatoma H22 is 33.61%, 40.16% and 55.74%. A total of 14 compounds were identified in the ethyl acetate fraction of Sauromatum giganteum (Engl.) Cusimano & Hett. The results of the experimental studies proved the antitumor activity of Sauromatum giganteum (Engl.) Cusimano & Hett and supported the traditional use of this plant. These data indicate the potential for the use of ethyl acetate fraction of Sauromatum giganteum (Engl.) Cusimano & Hett Tuber in tumor therapy, anti-tumor activity on cancer cell line in descending order is SGC-7901 > SMMC-7721 > MCF-7 > HT-29 > A549 > HeLa > MDA-MB-231.

Alkaloids from beach spider lily (Hymenocallis littoralis) induce apoptosis of HepG-2 cells by the fas-signaling pathway.

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose (0.8µg/ml) significantly inhibiting proliferation . The non- tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

Downregulation of Cdk1 and cyclinB1 expression contributes to oridonin-induced cell cycle arrest at G2/M phase and growth inhibition in SGC-7901 gastric cancer cells.

BACKGROUND: Oridonin isolated from Rabdosia rubescens, a plant used to treat cancer in Chinese folk medicine, is one of the most important antitumor active ingredients. Previous studies have shown that oridonin has anti- tumor activities in vivo and in vitro, but little is known about cell cycle effects of oridonin in gastric cancer. MATERIALS AND METHODS: MTT assay was adopted to detect the proliferation inhibition of SGC-7901 cells, the cell cycle was assessed by flow cytometry and protein expression by Western blotting. RESULTS: Oridonin could inhibit SGC-7901 cell proliferation, the IC50 being 15.6 µM, and blocked SGC-7901 cell cycling in the G2/M phase. The agent also decreased the protein expression of cyclinB1 and CDK1. CONCLUSIONS: Oridonin may inhibit SGC-7901 growth and block the cells in the G2/M phase by decreasing Cdk1 and cyclinB1 proteins.

Ethanol but not aqueous extracts of tubers of Sauromatum Giganteum(Engl.) Cusimano and Hett inhibit cancer cell proliferation.

BACKGROUND: Both alcohol and aqueous extracts of Sauromatum giganteum(Engl.) Cusimano and Hett, the dried root tuber of which is named Baifuzi in Chinese, have been used for folklore treatment of cancer in Northeast of China. However, little is known about which is most suitable to the cancer therapy. MATERIALS AND METHODS: Serum pharmacology and MTT assays were adopted to detect the effects of ethanol and aqueous extracts of Sauromatum giganteum(Engl.) Cusimano and Hett , prepared by heat reflux methods, on proliferation of different cancer cells. RESULTS: Cancer cells treated with medium supplemented with 10%, 20%, 40% serum(v/v) containing ethanol extract had a decline in viability, with inhibition rates of 7.69%, 21.8%, 41.9% in MCF-7 cells, 42.8%, 48.1%, 51.8% in SGC-7901 cells, 44.1%, 49.2%, 53.7% in SMMC-7721 cells, 6.8%, 15.2%, 39.8% in HepG2 cells, 7.57%, 16.3%, 36.2% in HeLa cells, 6.24%, 12.5%, 27.4% in A549 cells, and 7.20%, 17.5%, 31.3% in MDA-MB-231 cells, respectively. Viability in the aqueous extract groups was no different with that of controls. CONCLUSIONS: An ethanol extract of Sauromatum giganteum(Engl.) Cusimano and Hett inhibited the proliferation of SMMC-7721, SGC-7901 and MCF-7 cells, which supports the use of alcoholic but not aqueous extracts for control of sensive cancers, which might include hepatocarcinoma, gastric cancer and breast cancer.

[Study on the effects of two kinds of cactus polysaccharide on erythrocyte immune function of S180 mice].

OBJECTIVE: To study the effects of two kinds of cactus polysaccharide on erythrocyte immune function in S180 mice. METHOD: Classical pharmaceutical method and test kit. RESULT: The cactus polysaccharide increased the content of RBC-CaR, RFER, decreased the content of RFIR, raised the content of sialic acid. And the effect of median dose group of medical cactus polysaccharide and high dose group of edible cactus polysaccharide is very remarkable (P < 0.01) compared with model group. CONCLUSION: The cactus polysaccharide improved the erythrocyte function of tumor-mice, which may be one of anti-tumor mechanisms.

Effect of Haimiding on the functioning of red cell membrane of FC and H22 tumor-bearing mice.

AIM: To study the effect of Haimiding on the functioning of red cell membrane of FC and H(22) tumor-bearing mice. METHODS: The membrane fluidity of red cells is measured with DPH fluorescence probe as a marker; the amount of red cell membrane proteins is measured using polyacrylamide gel electrophoresis; the amount of sialic acid (SA) on the surface of red cell membrane and the sealability of these cells are measured using colorimetric analysis. RESULTS: Haimiding can lower the membrane fluidity of red cells in tumor-bearing mice and the amount of their membrane proteins, while increasing the amount of sialic acid in the membrane of red cells in these mice and enhancing the ability of the membrane of their red cells to reseal. CONCLUSION: The anti-tumor effect of Haimiding on tumor-bearing mice is due to its ability to improve and restore the functions of the membrane of their red cell and to enhance the immune effect of the organisms.

Anti-neoplastic efficacy of Haimiding on gastric carcinoma and its mechanisms.

AIM: To study the anti-neoplastic effect of Haimiding and its mechanisms of action. METHODS: Experiments using MTT and colony formation were carried out to study the in vitro anti-neoplastic action of Haimiding, its in vivo anti-neoplastic action was studied by observing its effect on the weight of tumors in FC mice and S(180), H(22) tumor bearing mice, as well as their life spans. The effect of Haimiding on cell apoptosis and different stages of cell cycles in human gastric carcinoma cells were studied by flow cytometry. Its effect on [Ca(2+)](i) of human gastric carcinoma cells and the source of Ca(2+) during the change of [Ca(2+)](i) were observed by confocal laser scanning technique. RESULTS: Haimiding showed a definite cytotoxicity to 8 human tumor cell lines, which was most prominent against BGC-823, E(ca-109) and HCT-8 tumor cells. It also exhibited an obvious inhibition on colony formation of the above tumor cell lines, which was most prominent in E(ca-109) tumor cells. It showed obvious inhibition on the growth of tumor in FC mice and S(180) bearing mice as well as prolonged the life span of H(22) bearing mice. It was able to induce apoptosis and elevate intracellular [Ca(2+)](i) concentration of tumor cells. The source of Ca(2+) came from both extracellular Ca(2+) influx and intracellular Ca(2+) release. CONCLUSION: Haimiding is composed of a TCM preparation and 5-flurouracil. Its anti-neoplastic potency is highly enhanced by synergism as compared with either one of its components. Its mechanisms of anti-neoplastic action can be attributed to its action to initiate apoptosis of tumor cells by opening the membrane calcium channel and inducing intracellular Ca(2+) release to elevate [Ca(2+)](i) of the tumor cells.

[Influence of sargassum fusiforme polysaccharide on apoptosis of tumor cells].

OBJECTIVE: To study mechanism of antitumor activity of Sargassum Fusiforme Polysaccharide (SFPS). METHOD: The effect on cell cycle and apoptosis was studied with flow cytometry (FCM). Intracellular calcium concentration [Ca2+]i was marked with Fluo-3/AM and measured with laser scanner confocal microscope (LSCM). RESULT: SFPS inhibited G0/G1 stage SGC-7901 from entering to S stage and increased APO%. The [Ca2+]i showed a transient rise and return to the original level. The concentration could be raised again by administering CaCl2. CONCLUSION: The antitumor effect of SFPS seems to be accomplished through the apoptosis associated with the increase in intracellular calcium concentration. Intracellular stores release the calcium during its action.

[Study on the effects of two kinds of cactus polysaccharide on erythrocyte membrane protein and fluidity of the lipid in S180 mice].

OBJECTIVE: To study the effects of two kinds of cactus polysaccharide on Band 3 protein, cross-linking protein and lipid fluidity of erythrocyte membrane in S180 mice. METHOD: The membrane protein content was analysed by SDS-PAGE. Lipid fluidity was measured by Skinitzky method. RESULT: The two kinds of cactus polysaccharide increased the content of Band 3 protein and decreased the content of cross-linking protein, raised the lipid fluidity. While the effect of median dose group of medical cactus polysaccharide is very remarkable (P < 0.01), the effect of high dose group of edible cactus polysaccharide is very remarkable (P < 0.01). CONCLUSION: By improving the erythrocyte membrane function of tumor-mice, they enhanced the immune function, which may be one of anti-tumor mechanisms.