Scientists have been studying the active ingredients of ganoderma lucidum, polysaccharides and triterpenoids are known as the key medicinal constituents. In recent 25 years, researchers over the world have studied the pharmacology of ganoderma lucidum extensively, especially the anti-tumor and immunomodulatory effects of ganoderma lucidum triterpenes and ganoderma lucidum polysaccharides.

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1 The Immunomodulatory Effects Of Ganoderma Lucidum
1.1 Cellular Immunity
The cellular immunity is major part of anti-tumor immunities, cellular immunity involves a variety of
immune cells. Ganoderma lucidum polysaccharides increased the cytoplasmic free Ca2 + concentration of peritoneal macrophages in mice significantly, in vitro ganoderma lucidum polysaccharides significantly promoted the protein kinase C activity of peritoneal macrophages
in mice.
Our experiments showed that ganoderma lucidum extract and ganoderma lucidum polysaccharide GL-B could significantly increase the secretion of tumor necrosis factor (TNF-α) and interferon-γ (IFN-γ) in a dose-dependent manner, and also confirmed that the anti-tumor effects of ganoderma lucidum polysaccharides were related to TNF-α and IFN-γ. Further researches on molecule level showed, ganoderma lucidum polysaccharides could promote the expression of TNF-α and IFN-γmRNA, which increased the transcription of TNF-α and IFN-γ mRNA.

Ganoderma lucidum polysaccharides could promote the expression of IL-12 P40 mRNA in dendritic cells , which increased the protein content of IL-12 P40 in culture supernatant. Further studies have shown that, after murine bone marrow-derived dendritic cells were sensitized by P815 tumor antigen, the specific cytotoxic T lymphocytes(CTL) induced by the DCs had the ability to kill P815. and ganoderma lucidum polysaccharides could enhance the ability.

This enhancement could be related to the ability of ganoderma lucidum polysaccharides to promote IFN-γ mRNA transcription and protein expression in CTL cells, which also improved the expression of granzyme B in CTLs in gene transcription and protein expression. Ganoderma lucidum polysaccharides improved the immununity functions of ehrlich ascites carcinoma and S180 sarcoma mice.

Ganoderma lucidum polysaccharides enhanced the immunomodulatory activity of immune effector cells in immunosuppressed mice.

A clinical study on patients suffering from terminal cancers has shown that the average concentration of IL-2, IL-6 and IFN-γ in plasma increased considerably after patients took Ganopoly (R), a polysaccharide component of ganoderma lucidum extract, and the concentration of TNF-α and IL-1 decreased significantly; meanwhile, NK cell activity was significantly increased. The study showed that ganoderma lucidum had an immunomodulatory effect on patients suffered from terminal cancers.

In another clinical study on patients suffered from terminal colon cancer showed that: ganoderma lucidum polysaccharides could increase the mitotic response of phytohaemagglutinin(PHA) and increase the
concentration of IL-2, IL-6 and IFN-γ in plasma to enhance NK cell activity, but the concentration of TNF-α and IL-1 decreased.

In vitro the immune protein LZ-8 in ganoderma lucidum had a strong mitogenic effect on spleen cells in mice. In vivo ganoderic acid Me promoted the expression of IL-2, IFN-γ and NK cell activity. Ganoderma lucidum polysaccharides took the main role of immunomodulator, while ganoderma lucidum triterpenoids also enhanced NK cell activity and improved the level of cytokine expression.

1.2 Humoral Immune Function
Ganoderma lucidum also showed significant humoral immune activity. The SRBC-induced PFC reaction in normal mice could be greatly improved by intraperitoneal injection(1mg/kg and 5mg/kg) of ganoderma lucidum polysaccharides BN3 in IP mice, which proved that ganoderma lucidum had the ability to promote the humoral immune functions in normal mice.

Related scientific experiments showed that fructose from ganoderma lucidum stimulated lymphocyte proliferation in mice, B cells increased 3-4 times, and activated mouse lymphocytes to express CD71 and CD25, which indicated immunoglobulin secretion increased,too.

In vitro two heteropolysaccharides and one glucan from ganoderma lucidum stimulated the proliferation of T- and B-lymphocytes in a dose-dependent manner. Ganoderma lucidum polysaccharides could increase the number of immune cells by promoting the proliferation and differentiation of immune cells in tumor-bearing mice. In recent years, Peking University Medical Department studies have shown that membrane Ig and TLR-4 were the immune receptors of B cells activated by ganoderma lucidum polysaccharides(GLPS), TLR-4 was related to the activation of
macrophages by GLPS.

In summary, ganoderma lucidum had cell and humoral immunomodulatory effects. From the immunological point of view, the anti-tumor effects of ganoderma lucidum and its activation of immune effector cells (especially macrophages and spleen cells), promoted the expression of T cells in
TNF-α and IFN-γ mRNA in macrophages.

2 The Antitumor Effect Of Ganoderma Lucidum
2.1 In Vitro Anti-tumor Study

The triterpenoids from ganoderma lucidum had obvious cytotoxicity in vitro. The triterpenoids, ganoderal A and dihydroganoderal A, had strong antitumor activities in vitro. Studies showed that the ED50 values of ganoderic acid γ, ε and θ on sarcoma cell lines Meth-A were less than 15μg/mL. The ED50 value of ganoderic acid θ was 5.7μg / mL; the values on lewis lung carcinoma(LLC) were not as good as expected, only the ED50 value of ganoderic acid θ was less than 15μg/mL. Lucidenic acid N, lucidenic acid A and ganoderic acid E also had strong cytotoxic
effects on liver cancer Hep G2 and P-388 tumor cells.

In vitro anti-cancer experiments showed that lucialdehydes B, C, ganodermanonol and ganodermanondiol had a strong cytotoxic effect on tumor cells, in which the of effect of lucialdehydes C was the strongest, its ED50 values of LLC, T-47D, S180 and Meth-A were 10.7, 4.7, 7.1 and 3.8 μg/mL respectively. Ganoderma lucidum extract could inhibit the proliferation of MCF-7 human breast cancer cells in a dose and time-dependent manner, and induce MCF-7 apoptosis.

Researchers found that the constituent WEES-G6(which is rich in ganoderic acids) from ganoderma lucidum inhibited the growth of Huh-7 human hepatoma cells, but did not inhibit the growth of normal liver cell lines. In vitro ganoderic acids significantly inhibited the growth of human hepatoma cell line BEL7402 . Our studies showed that triterpenoids from ganoderma lucidum inhibited the proliferation of sarcoma cell L290, intestinal cancer cell SW620, K562 leukemia cell and BEL7402 liver cancer cell.

Ganoderma lucidum could induce tumor cell differentiation. Ganoderma lucidum extract inhibited the proliferation of mouse pheochromocytoma cells (PC12) and promoted their differentiation. After being treated with ganoderma lucidum extract, the morphology of PC12 cells changed significantly.

In addition, ganoderma lucidum extract induced the expression of neuronal marker protein in PC12, including two phosphorylated neurofilament proteins with 200kDa and 160kDa size respectively, which were the main structural proteins of nerve cytoskeleton. This indicated that there were some substances in ganoderma lucidum extract that induced PC12 tumor cells. Further studies have shown that ganoderma lucidum extract could activate extracellular regulated kinase 1 (Erk1) and Erk2 as well as cAMP-response element binding protein (CREB).

In addition, the water-soluble constituents of ganoderma lucidum mycelia culture medium also induced the differentiation of leukemia cells HL-60. The differentiation rate of HL-60 cells was significantly increased after 4-day treatment. The induction of cell differentiation was one of the mechanisms of ganoderma lucidum anti-tumor effects.

Ganoderic acids induced tumor cell apoptosis. Ganoderic acids could inhibit the growth and induce the apoptosis of 95-D cells in vitro. Ganoderma lucidum could inhibit the transfer of breast cancer and prostate cancer cells. Ganoderma lucidum inhibited the constitutively active transcription factors AP-1 and NF-B in breast cancer MDA-MB-231 and prostate cancer PC-3 cells (both uPA and uPAR containing AP-1 and NF -B DNA
binding sequence), this was because ganoderma lucidum could reduce AP-1 and NF-.B constitutive activation, and ultimately the uPA and uPAR expression and uPA secretion (in the two cancer cells) were inhibited.

The study showed that ganoderma lucidum inhibited the transfer of cancer cells by the inhibition of uPA and uPAR transcription factor AP-1 and NF-.B, uPA and uPAR expression and uPA secretion were inhibited, and the transfer of breast and prostate cancer cells were inhibited. The preliminary study revealed the molecular mechanism of ganoderma lucidum to inhibit the transfer of breast and prostate cancer cells.

2.2 In Vivo Anti-tumor Study
Ganoderma lucidum polysaccharides had significant anti-tumor effects in vivo. The monosaccharide components of polysaccharides AS-1 included mannose 67.5%, xylose 22.5%, glucose 5.8%, galactose 1.8% and ribose 2.0%. %. The antitumor activity of AS-1 on S-180 was up to 97.5%. Two polysaccharides with relative molecular weights of 1,050,000 and 450,000 from ganoderma lucidum could inhibit the growth of S-180 in ICR-Jcl mice. The 50% inhibitory doses were 8.0mg/Kg and 22.0mg/kg.

Proteoglycans from mycelia of ganoderma lucidum significantly prolonged the lifespan of mice bearing lymphoblastic leukemia L1210 cells. Ganoderma lucidum polysaccharides could inhibit the growth and metastasis of fibrosarcoma in C3H mice and prolong the lifespan of Lewislungcarcinoma tumor-bearing mice.

In addition, the mycelial extract of ganoderma lucidum had a significant inhibitory effect on malignant muscle fiber tumor in C3H mice, which was also effective on metastatic pulmonary lesions.

Ganoderma lucidum triterpenoids also showed significant antitumor activities in vivo. Ganoderic acids F inhibited the growth of early transplanted Lewislungcarcinoma (LLC) tumor and inhibited its transfer to the liver and the growth of metastatic tumor. And ganoderic acids F inhibited the tumor angiogenesis induced by matrigel, vascular endothelial growth factor (VEGF) and heparin in vitro. Both ganoderma triterpenoids and acidic triterpenes inhibited the growth of B16 melanoma cells in C57BL/6 mice. The triterpenoids had stronger inhibitory
effects.

2.3 The Antitumor Merchanism Of Ganoderma Lucidum
The mechanism of anti-tumor effects of ganoderma lucidum has been the centre of attention. As the chemical constituents of ganoderma lucidum were complex, which could inhibit many types of tumors, the mechanism of their anti-tumor effects was inconclusive.

It was known that the immune-mediated effect of the host was one of the main mechanism of anti-tumor of ganoderma lucidum polysaccharides. Professor Lin Zhibin, Department of Medicine of Peking University, has achieved remarkable achievements in this direction. His recent study found that membrane Ig and TLR-4 were the immune receptors by which ganoderma lucidum polysaccharides activated the B cells, and TLR-4 was also related with macrophages activation by ganoderma lucidum polysaccharides.

In addition, the other possible merchanism of ganoderma lucidum polysaccharides to prevent and treat cancers included antioxidation, induced cell differentiation and activation of mitogen-activated protein (MAP) kinase and
inhibition of tumor angiogenesis. The anti-cancer mechanism of ganoderma lucidum triterpenoids included antioxidation, induced II phase enzyme, direct cytotoxicity, inhibition of tumor angiogenesis, and regulation of signal transduction. Ganoderma lucidum spores or fruiting bodies also inhibited urokinase plasminogen activator (uPA) and its receptor (uPAR) and so on.

In addition, ganoderma lucidum polysaccharides significantly inhibited the adhesion of malignant human breast tumor cells and integrin expression. Further studies have shown that ganoderma lucidum polysaccharides could significantly reduce the expression of cell adhesion molecule β1 integrin. As the adhesion of tumor cells was of great importance to tumor cell formation, growth and metastasis, therefore, ganoderma lucidum polysaccharides inhibited tumor cell adhesion and the corresponding integrin expression, helped us to understand the anti-tumor mechanism of ganoderma lucidum polysaccharides.

Discussion
Ganoderma lucidum polysaccharides take the immunoregulatory effects mainly. Ganoderma lucidum polysaccharides significantly enhance the immunity, such as enhancing delayed hypersensitivity in mice, promoting lymphocyte proliferation response, improving cytotoxic functions, macrophage phagocytosis, NK cell cytotoxicity and cytokine production; and ganoderma lucidum polysaccharides and triterpenoids take the anti-tumor effects, it is not clear if synergistic reactions occur between both components. Ganoderma lucidum triterpenoids have direct cytotoxic effects, Ganoderma lucidum polysaccharides do not have the ability to kill tumor cells in vitro.

In terms of anti-tumor mechanism of ganoderma lucidum, it is generally believed that the key of anti-turmor is to enhance the body immunity and activate the immune effector cells to release cytokines and kill tumor cells.
As tumor formation is a complex multi-stage process, the tumor can be prevented or cured by blocking any of the stages. Some active substances of ganoderma lucidum can block one or some of the stages.

For example, ganoderma lucidum polysaccharides can significantly improve human immune system and prevent tumor formation through the host immune mediation; ganoderma lucidum polysaccharides or triterpenoids can enhance the detoxification activity by inducing phase II enzymes.

Ganoderma lucidum polysaccharides can inhibit the adhesion of tumor cells and the expression of the corresponding integrin, and ganoderma lucidum polysaccharides can inhibit the proliferation of tumor cells by inhibiting the tumor angiogenesis. Ganoderma lucidum spores or fruiting bodies can also reduce the invasiveness of cancer cells or inhibit their metastasis by inhibiting uPA and its receptor uPAR.