Progress on association between autophagy and cancer / 浙江大学学报·医学版

Autophagy is an evolutionarily well conserved cellular catabolic process.Cellular proteins and organelles are engulfed to autophagosomes and eventually delivered to lysosomes for degradation. Autophagy is closely associated with a variety of human diseases especially cancer. At present, it has been widely accepted that autophagy is a "double-edged sword" in cancer: it blocks tumorigenesis at the early stage, while it facilitates tumor development at the promotion and progression stage. Moreover, autophagy can be induced by chemotherapy and radiotherapy, which is generally play a pro-survival function. Thus, autophagy is an important target for cancer therapy, and suppression of autophagy to enhance the efficacy of cancer therapeutic agents is a novel strategy in cancer therapy under active clinical trials.

Effect of Achyranthes bidentata polysaccharides stimulated dendritic cells co-cultured with cytokine induced killer cells against SW480 cells / 中国中药杂志

Achyranthes bidentata polysaccharides (ABPS) was extracted from the root of A. bidentata. Dendritic cells (DC), which were stimulated with ABPS and/or tumor antigen SW480, were co-cultured with cytokine induced killer cells (CIK) to test the cytotoxic effect on colon cancer cell line SW480. Peripheral blood mononuclear cells (PBMNCs) which were separated from human peripheral blood were cultured to DC and CIK separately. (1) DC were divided into four groups: pure DC served as control group; ABPS (50 mg x L(-1)) stimulated DC served as experimental group; SW480 tumor antigen stimulated DC served as the second experimental group; ABPS (50 mg x L(-1)) and SW480 tumor antigen co-stimulated DC served as the third experimental group. Flow cytometry was used to detect the difference of the positive rate of molecules in the cell surface of DC, include CD80, CD86, CD1c, CD40, HLA-DR (6 samples for each group). (2) The four DC groups were mixed with CIK at the ratio 1:5 and acted as effect cells (DC + CIK groups), and the colon cancer cell line SW480 acted as target cells. The effect cells and the target cells were mixed together at the ratio 30: 1, 20:1 and 10:1 separately, and the CCK-8 kit was used to test the cytotoxic effect on colon cancer cell line SW480. (3) At the mixing ratio 30:1 of effect cells and target cells, ELISA was used to test the level of cytokines secretion, including IL-2, IL-12p70, IL-17 and TNF-alpha, in the liquid supernatant of every test group (3 duplication per sample). The results showed as following: (1) The positive rates of CD80, CD11c, HLA-DR, in the cell surface of DC which was co-stimulated by ABPS (50 mg x L(-1)) and SW480 tumor antigen, were obviously higher than the other DC groups (P < 0.05), and the positive rates of CD86, CD40 were obviously higher than the pure DC group (P < 0.05), and there was no remarkable difference with the other two DC groups. (2) At the mixing ratio 30:1, 20:1 and 10:1 of the effect cells and the target cells, the cytotoxic effect of ABPS stimulated DC + CIK group and SW480 tumor antigen stimulated DC + CIK group was obviously higher than DC + CIK group (P < 0.05), the cytotoxic effect of ABPS and SW480 tumor antigen co-stimulated DC + CIK group was obviously higher than all the other groups. (3) At the mixing ratio 30:1 of the effect cells and the target cells, the secretion levels of IL-12p70 and TNF-alpha in the liquid supernatant of the ABPS and SW480 tumor antigen co-stimulated DC + CIK group were obviously higher than all the other groups (P < 0.05), the secretion levels of IL-2 and IL-17 in the liquid supernatant of every test group have no remarkable difference. The cytotoxic effect of ABPS stimulated DC + CIK on SW480 was obviously increased. The cytotoxic effect of ABPS and SW480 tumor antigen co-stimulated DC + CIK group was obviously higher than all the other.

Preventive effect of IL-18 gene modified mature dendritic cells vaccine on airway inflammation in mouse asthma model / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the preventive effect of interleukin-18 (IL-18) gene modified mature dendritic cells (mDC) vaccine on airway inflammation in mouse asthma model.</p><p><b>METHODS</b>The asthma model was induced by injection of ovalbumin (OVA) in BALB/c mice. IL-18 gene modified mouse mature dendritic cells (mDC) were detected by flow cytometry and its capacity of inducing allogeneic T cell responses was examined by mixed lymphocyte reaction (MLR). The OVA-induced asthmatic mice were randomly divided into 6 groups: PBS group, DXM group, mDC group, Ad-LacZ-mDC group, Ad-IL-18-mDC group and control group. The pathological changes in lung tissues were assayed by HE and AB-PAS staining. The numbers of inflammatory cells and percentage of eosinophils (EOS) in bronchoalveolar lavage fluid (BALF) were counted. The levels of IFN-γ IL-4 and IL-13 in culture supernatant of splenocytes were measured by ELISA method. The percentage of CD4(+)CD25(+)Foxp3(+) Treg was assessed by flow cytometry analysis.</p><p><b>RESULT</b>The vaccine was effective in decreasing the infiltration of EOS and accumulation of airway goblet cells in lung tissues, the numbers of inflammatory cells and percentage of EOS in BALF, and the levels of IL-4 and IL-13 in culture supernatant of splenocytes, and in increasing the levels of IFN-γ in culture supernatant of splenocytes and the percentage of CD4(+)CD25(+)foxP3(+) reg.</p><p><b>CONCLUSION</b>IL-18 gene modified mDC vaccine has a preventive effect on airway inflammation in OVA-induced asthmatic mice.</p>

Promoting effect on NK cells induced by myeloid-derived suppressor cells in acute fulminate hepatitis of mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the interaction between myeloid-derived suppressor cells (MDSCs) and nature killer cells during acute fulminate hepatitis.</p><p><b>METHODS</b>Acute fulminate hepatitis were induced by i.p. co-injection of LPS and D-GalN in mice, and the ratio of MDSCs,NK cells and the activation of NK cells in different tissues were analyzed by FACS at 0 h,1.5 h,3 h and 6 h.</p><p><b>RESULTS</b>The percentage of MDSCs and NKG2D+NK cells in different tissues increased as acute fulminate hepatitis progressed, with the increased NK cells in liver tissue. The mean fluorescence intensity of NKG2D on NK cells in different tissues were also enhanced.</p><p><b>CONCLUSION</b>MDSCs induce the proliferation and activation of NK cells in mice with acute fulminate hepatitis.</p>

Preparation of anticolon carcinoma vaccine with rich chaperone peptides and study on its anticancer efficacy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy.</p><p><b>METHODS</b>CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50,000 and above 300,000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70,000, 90,000, 95,000, 110,000 and 170,000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL.</p><p><b>RESULTS</b>Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK(P<0.01).</p><p><b>CONCLUSIONS</b>The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42 centi-degree heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.</p>

Effect of macrocalyxin A on HL-60 cells proliferation, differentiation and apoptosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of macrocalyxin A (MA) on proliferation, differentiation and apoptosis in HL-60 cells and explore its possible mechanisms.</p><p><b>METHODS</b>Different concentration of MA were used to treat HL-60 cells. Proliferation inhibition was analyzed by Trypan blue staining and MTT assay, cell apoptosis by cell morphology, DNA content, cell cycle analysis, Annexin-V/PI and Hoechst 33258 fluorescence staining. The differentiation of HL-60 cells was evaluated by cell morphology, NBT tests and expression of CD11b, CD13, CD14. The expressions of bcl-2, bax, Fas, P53, mitochondrial transmembrane-potential (DeltaPsim) and mitochondrial membrane protein were analyzed by flow cytometry.</p><p><b>RESULTS</b>MA could inhibit HL-60 cells proliferation capacity in a time-and dose-effect, with a 24 h IC50 value of 8.76 microg/ml, 48 h of 7.17 microg/ml and 72 h of 7.14 microg/ml. The HL-60 cells apoptosis was confirmed by cell morphology, sub-G1 phase and Annexin-V/PI labeling in a time and dose dependent manner. The more mature HL-60 cells were a with higher positivity of NBT and expressions of CD11b than those cultured without MA. The expression of bax was increased, while bcl-2, P53, Fas were unchanged on the MA treatment. MA could increase the expression of mitochondrial membrane protein in a dose-dependent manner while the DeltaPsim was reduced.</p><p><b>CONCLUSION</b>MA can inhibit proliferation and induce differentiation and apoptosis of the HL-60 cells. The mechanism may be related with up-regulating bax, opening the mitochondrial permeability transition pore and reducing DeltaPsim.</p>

Surface markers and functions of human dendritic cells exposed to mobile phone 1800 MHz electromagnetic fields / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of mobile phone 1800 MHz electromagnetic fields (EMF) on the surface markers and the functions of human dendritic cells (DC).</p><p><b>METHODS</b>Human DCs were exposed to intermittent 5 min on/10 min off EMF with specific absorption rates (SAR) 4 W/kg for 0 h, 1 h, 12 h or 24 h, respectively. FACS analysis was used to detect the positive percentage of DC surface markers including HLA-DR and co-stimulatory molecules such as CD80, CD86, CD40 and CD11c. CCK-8 kit was adopted to examine the function of allo-mixed lymphocyte reaction (allo-MLR) of DC, and enzyme linked immunosorbent assay (ELISA) to identify the levels of IL-12p70 and TNF-alpha secreted by DC.</p><p><b>RESULT</b>Compared with the sham radiation group, after exposure to the electromagnetic fields for 1 h, 12 h, or 24 h, HLA-DR, CD80,CD86 and CD40 were all declined except CD11c. The ability of DC allo-MLR in each exposure group was decreased significantly (P<0.05), especially in the 24 h exposure group. However, the secreted levels of IL-12p70 and TNF-alpha of DC in each exposure group remained no changed.</p><p><b>CONCLUSION</b>The study showed that EMF exposure could down-regulate the surface molecules and stimulation ability of human DC.</p>

Study on effect of lentinan in enhancing anti-tumor action of dendritic cytoma vaccine and its mechanism / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To improve the anti-tumor effect of dendritic cytoma vaccine (DCV) for finding an effective anti-tumor biotherapy.</p><p><b>METHODS</b>DC vaccine prepared by transfection of adenovirus mediated melanoma-associated antigen gene (gp100) into bone marrow-derived dendritic cell (DC) was used to study the immuno-therapeutic effect and the mechanism of lentinan (LNT) in different dosages, used alone or combined with gp100-DC for treatment of B16 melanoma bearing mice.</p><p><b>RESULTS</b>After being treated with LNT combining gp100-DC, the growth of malignant melanoma was inhibited with the tumor-free survival in the experimental animals being 66.7%. The treatment could also significantly enhance the activity of cytotoxicity T lymphocyte (CTL) and natural killer (NK) cells, elevate the levels of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) in splenocytes, and histological examination showed that a large amount of inflammatory cells infiltrated inside and around the tumor, and obvious necrosis of tumor cells was found.</p><p><b>CONCLUSION</b>By combined use with LNT the anti-tumor immuno-reaction of DCV vaccine could be enhanced effectively.</p>

Serum levels of sFas, sICAM-1, IL-18 in patients with chronic hepatitis C and their clinical significance / 浙江大学学报·医学版

OBJECTIVE: To investigate the serum levels of soluble Fas antigen (sFas), soluble intercellular adhesion molecules-1 (sICAM-1), interleukin-18 (IL-18) in patients with chronic hepatitis C and to study their roles in pathogenesis of chronic hepatitis C. METHODS: Serum sFas, sICAM-1, IL-18 levels were measured in 30 cases of chronic hepatitis C before and after treatment of interferon-alpha by enzyme-linked immunosorbent assay (ELISA), serum titer of HCV-RNA was detected by quantitative PCR and serum ALT activity was also detected. RESULTS: Serum levels of sFas sICAM-1 IL-18 in chronic hepatitis C patients were significantly higher than those in normal controls (P<0.01), showing correlation with serum HCV-RNA titer (r=0.915, r=0.795, r=0.757, respectively, P<0.01), Serum levels of sICAM-1, IL-18 showed correlation with serum ALT level(gamma=0.952, gamma=0.969, respectively, P<0.01), but no relationship was observed between serum sFas and serum ALT level(P>0.05). Serum levels of sFsa sICAM-1 IL-18 markedly decreased in responsive patients while no change was observed in patients with no response after treatment. CONCLUSION: Soluble Fas, soluble ICAM-1, IL-18 may participate in the pathogenesis of chronic hepatitis C and show correlation with the severity of histological inflammation and viral titer.