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Objective To investigate the effects of β adrenergic receptor blocker carvedilol and cholinergic receptor blocker scopolamine on the biological behaviors of prostate cancer in C57BL/6 mice.Methods From June 2014 to November 2014, sixty male C57BL/6 mice, 10 weeks old, were injected with RM-1 cells (0.5 × 106) into bilateral dorsal prostate capsules of each mice by microsyringe and were randomly divided into three groups, including carvedilol group (K), scopolamine group (D) and saline group (N) after modeling.In K group, the carvedilol (10 mg/kg) was given to mice through gastric tube.The normal saline was given to them by subcutaneous injection.In D group, the normal saline was given to mice through gastric tube.Meanwhile, the scopolamine (1 mg/kg) was injected subcutaneously.In N group, daily normal saline was given by through gastric tube and subcutaneous injection.At the seventh day of postoperation, five mouse were sacrificed in each group every three days to observe the local growth,invasion and metastasis of prostate cancer to pelvic nodes and liver.Immunohistochemical determination of prostate tumor was made by TH staining and VAChT staining in each group.Results The volume of the prostate cancer was gradually increased in three groups.There was no statistical difference of prostate volume among three groups on the day 7 and 10 post-operatively (all P ＞ 0.05).The prostate volume of K group, D group and N group on the day 13 were 0.05 ± 0.04, 0.18 ± 0.08, 0.14 ± 0.05 cm3, respectively (P ＜0.05).However, the statistical significance existed between K and N group (P ＜ 0.05).There was no statistical difference of AOD between TH and VAChT among three groups on the day 7 (all P ＞ 0.05).The AOD of TH and VAChT in prostate cancer of K group, D group and N group on the day 10 were [(0.0114±0.016), (0.114±0.002), (0.059±0.008)] and [(0.025 ±0.011), (0.0226±0.003), (0.009±0.003)], respectively (P ＜0.05).Those values on the day 13 were [(0.147 ±0.036),(0.129 ±0.025), (0.071 ±0.022)] and [(0.020 ±0.005), (0.020 ±0.002), (0.010 ±0.002)],respectively (P ＜0.05).TH and VAChT expression of K and D in 10th and 13th is higher than in N (all P ＜ 0.05).At the 7th day, no metastasis of pelvic nodes was detected in all groups.At 10th day, 2 cases of K, no case of D and 3 cases of N were detected the lymph node metastasis.At the 13th day, 3 cases of K, 1 case of D and 4 cases of N were recorded the lymph node metastasis.At the 7th day, no metastasis of liver was detected.At 10th day, 1 case of K, no case of D and 2 cases of N were found the liver metastasis.At the 13th day, 3 cases of K, 1 case of D and 4 cases of N were recorded the liver metastasis.The lifesapn of tumor-bearing mice in K group, D group and N group was 16.8 ±0.8, 17.6 ±0.5, 15.8 ±0.1, respectively (P ＞ 0.05).Conclusions β adrenergic receptor blocker carvedilol can suppress the growth of prostate cancer and cholinergic receptor blocker scopolamine may can inhibit metastasis of prostate cancer.
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Mesenchymal stem cells (MSCs)have inherent tumor-trophic migratory properties and low immunogenicity,which allow them to serve as vehicles for delivering effective,targeted therapy to tumors.MSC plays an anti-tumor role by releasing some cytokines,which can be strengthened by oncolytic virus,antiangio-genesis agent,tumor necrosis factor guided apoptosis ligand,IL,IFN and pro-drug.In addition,there are some advantages in combination treatment of MSC and others therapies.
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<p><b>OBJECTIVE</b>To compare the immune function after laparoscopic surgery (LS) and conventional open surgery(OS) for colorectal cancer (CRC).</p><p><b>METHODS</b>PubMed, EMBASE, the Cochrane Library, China National Knowledge Infrastructure (CNKI), and Wanfang Database were systematically searched for randomized controlled trials published before August 2013 concerning the immunological difference between LS and OS. Data extraction was performed independently by two reviewers and data analysis was performed using Review Manager ver. 4.3.1.</p><p><b>RESULTS</b>Twelve studies including 638 patients (307 in LS group and 331 in OS group) were eligible for analysis. Overall analysis demonstrated that no significant differences were identified for blood C-reactive protein level on postoperative days (POD) 0-1 (P=0.40), plasma lymphocyte count on POD 1-3 (P=0.92) and POD 4-7 (P=0.64), plasma CD4⁺ T cell count on POD 1-7 (P=0.63), plasma CD8⁺ T cell count on POD 4-7 (P=0.09), and plasma NK cell count POD 1-3 (P=0.34) as well as POD 4-7 (P=0.46). Data analysis also showed that a significantly lower serum level of IL-6 on POD 0-1 after LS (WMD=-25.03, 95% CI:-34.06 to -15.99, P=0.000), and a significantly higher plasma level of CD8⁺ T cell count on POD 1-3 after LS(WMD=0.05, 95% CI:0.01 to 0.08, P=0.004).</p><p><b>CONCLUSIONS</b>Although postoperatively short-term humoral immune function trends to be better after LS for CRC compared to OS, there is no sufficient evidence to support superior preservation of global immune function after acute reactive phase.</p>
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Humans , Colorectal Neoplasms , Allergy and Immunology , General Surgery , Laparoscopy , Laparotomy , Randomized Controlled Trials as TopicABSTRACT
Aim To investigate the effects of PLCε on the invasion and migration of human osteosarcoma cancer cells U2OS. Methods RNA interference ( RNAi) was used to inhibit PLCεexpression, and the proliferation of cancer cells was measured by CCK-8 assay. The migration of the cells was measured by scratch wound healing assay and migration chamber as-say. Gelatin zymography was performed to measure the MMP2 activities in U2OS cells. Results PLCε ex-pression was suppressed by siRNA. CCK-8 assay showed that PLCε had no effect on the proliferation of cancer cells. PLCε knockdown inhibited cell invasion and activities of MMP2 . Conclusion PLCε knock-down can inhibit the migration and invasion of human osteosarcoma cancer cells U2OS.
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ObjectiveTo investigate the inhibitory effect of dacarbazine and an oncolytic adenovirus carrying interleukin-24 (IL-24) on transplanted melanoma in nude mice.MethodsNude mice were inoculated with human A375 melanoma cells to establish a model of malignant melanoma.Then,the mice were divided into 4 groups to be treated with an oncolytic adenovirus carrying interleukin-24 (ZD55-IL-24),dacarbazine,the combination of ZD55-IL-24 and dacarbazine,and phosphate buffer(PBS),respectively,for 3 days.Seven days after the end of the treatment,some mice were sacrificed followed by the determination of IL-24 and E1A protein levels in tumor tissue by Western blot.The tumor volume was measured on a daily basis for 30 days.ResultsIL-24 and E1A were highly expressed in melanoma cell-bearing nude mice treated with ZD55-IL-24 and dacarbazine.At 30 days after the inoculation,the average volume of transplanted melanoma was (2346.5 ± 576.0) mm3 in the combination group,significantly different from that in the ZD55-IL-24 group((4141.6 ± 1348.2) mm3,P ＜ 0.05),dacarbazine group((5230.1 ± 922.8) mm3,P ＜ 0.05),and the control group ((7135.1 ± 1002.3) mm3,P ＜ 0.05).ConclusionThe ZD55-IL-24 in combination with dacarbazine exhibits a remarkably inhibitory effect on the proliferation of melanoma transplanted into nude mice.
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Objective To investigate the expression of bone morphogenetic protein-7(BMP-7)in the tissue of prostate cancer(PCa). Methods The pathological samples of 87 cases of PCa were collected.The average age was 66(59-78)years,preoperative of t-PSA was 45.7(2.4-138.2)ng/ml.Gleason score:37 cases were≤6,18 cases were 7,32 cases were≥8.Stages:stage I(T1aN0M0)+stageⅡ(T1bN0M0,T1cN0M0,T2N0M0)20 cases;StageⅢ(T3N0M0)20;Stage Ⅳ(T4N0 M0,TxN1 M0,TxN0 M1)47 cases.According to bone scan or positron emission computed tomography-CT test results,patients were divided into PCa without bone metastasis,42 cases and PCa with bone metastasis,45 cases.Thirty cases of BPH were set as controls.BMP-7 in the PCa and BPH were detected by PV immunohistochemical study.Statistical analysis was done between two groups to compare the differential expression of BMP-7 and serum t-PSA in PCa, and BPH tissues.Results BMP-7 expression in the absorbance A value in benign prostatic hyperplasia was 70.55±5.41, in prostate cancer tissue 70.47± 6.18, no significant differences between the 2 groups(P>0.05).BMP-7 expression in the absorbance A value in prostate cancer without bone metastasis was 65.94 ± 1.76, but with bone metastasis 74.80±5.76.There was a significant difference (P<0.05).Gleason score≤6 absorbance A value was 65.96 ± 1.56, Gleason 7 absorbance A value 65.83 ± 2.75,≥8 absorbance A value 78.06±1.39.Compared with Gleason score≥8, BMP-7 expression in the absorbance A value were significantly lower than the latter (P<0.05).Clinical stage grouping of BMP-7 expression in the absorbance A value: Stage Ⅰ + Ⅱ 65.86±1.72, Stage Ⅲ 65.87±1.85, Stage 74.49±5.83.There was a significant difference (P<0.05).In PCa tissues, BMP-7 of the absorbance A value and the serum t-PSA values showed a positive correlation (r=0.77,P,<0.05). Conelusions The expression level of BMP-7 has occurred in the high pathological Gleason score, late clinical stage, particularly in bone metastasis cases.The expression level of BMP-7 and serum t-PSA have a positive correlation.
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Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene (ZD55-IL-24) on the apoptosis of human melanoma cell line A375. Methods The oncolytie adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-1L-24, ZD55 fused with enhanced green fluorescent protein (ZD55-EGFP), and replication-deficient adenovirus ZD55 carrying IL-24 gene (AD-IL-24), respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively.Subsequently, the eytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium (MTT) assay, respectively. The expressions of EIA and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirns contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious eytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time-and concentration-dependent manner. As shown by Western blot analysis, ZD55-1L-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.
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Objective To evaluate the antitumor effect of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice.Methods BALB/c nude mice were subcutaneously inoculated with A375 cells to establish a model of malignant melanoma.When the volume of implanted tumor reached 100-150 mm3,murine models were randomly divided into three groups to receive a 3-day intratumoral injection of IL-18 gene-expressing human oncolytic adenovirus named ZD55-IL-18,IL-18 gene-expressing adenovirus named Ad-IL-18,phosphate buffer saline(PBS),respectively.The tumor size was measured at an interval of 4 days for 9 weeks.Hematoxylin and eosin (HE)staining was performed to observe the morphological changes of tumor cells.The protein expression of IL-18 and E1A.microvessel density in tumor tissue,and apoptosis of tumor xenografts were detected by immuno fluorescence assay,immunohistochemistry and in situ end labeling technique (TUNEL).respectively.Results The treatment with ZD55-IL-18 significantly inhibited the growth of tumor.Forty-four days after the treatment,the mean tumor volume was 1039.378±29.67 mm3 in ZD55-IL-18-treated mice.significantly smaller than that in Ad-IL-18 treated mice(2900.46±62.65 mm3)and PBS-treated mice(3980.24±63.78 mm3).HE staining showed that the nuclei of tumor cells were heavily stained with few nucleoli in ZD55-IL-18-treated mice.Increased positivity rate of IL-18 was noticed in ZD55-IL-18-treated mice vs.AD55-IL-18-treated mice(83.4%±3.2%vs 24.4%±2.1%.P＜0.01).Moreover,immunofluorescence assay revealed the presence of E1A protein in tumor tissue.A decrease was found in the microvessel density in ZD55-IL-18-treated mice compared with the PBS-treated mice(P＜0.01).The apoptosis rate in tumor cells from high to low was 86.28%±3.25%in ZD55-IL-18-treated mice,43.67%±3.46%in Ad-IL-18-treated mice,and 10.73%±2.48%in PBS-treated mice;there was a significant difference between the three groups(all P＜0.05).Conclusion The oncolytic adenovirus expressing human IL-18 gene,ZD55-IL-18,has a significant inhibitory effect on the growth and metastasis of malignant melanoma implanted in nude mice.
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To investigate the effects of anti-sense peptide nucleic acids (PNAs) targeting Ki-67gene on modulation of the proliferation and apoptosis of human renal carcinoma cell lines, human renal carcinoma cell line 786-0 cells were treated with anti-sense PNAs at different concentrations (1.0 μmol/L, 2.0 μmol/L, 10.0 μmol/L). The Ki-67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-thymidine incorporation. The apoptosis of 786-0 cells was detected by TUNEL assay. The control groups were treated with anti-sense oligonucleotide (ASODNs)targeting Ki-67 gene. Our results showed that the Ki-67 expression of 786-0 cells treated with anti-sense PNAs (16.9±0.7) was significantly inhibited as compared with that of the control groups (28.6±0.4) (P＜0.01). The Ki-67 protein rate of 786-0 cells treated with anti-sense PNAs (42.1±2.2)was significantly reduced when compared with that of the control groups (83.6±1.4) (P＜0.01). Proliferation of 786-0 cells treated with anti-sense PNAs (20.7±1.5) was significantly inhibited as compared with that of the control groups (58.6±1.4) (P＜0.01). The apoptosis rate of 786-0 cells treated with anti-sense PNAs (28.7±2.3) was significantly increased higher compared with that of the control groups (13.8±1.0) (P＜0.01). From these finds we are led to conclude that anti-sense PNAs targeting Ki-67 gene have stronger effects on the inhibition of the proliferation and induction of apoptosis of human renal carcinoma cells than ASODNs targeting Ki-67 gene. The strategies using anti-sense PNAs targeting Ki-67 gene may be a promising approach for the treatment of renal cell carcinoma.
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To investigate the effects of anti-sense peptide nucleic acids (PNAs) targeting Ki-67 gene on modulation of the proliferation and apoptosis of human renal carcinoma cell lines, human renal carcinoma cell line 786-0 cells were treated with anti-sense PNAs at different concentrations (1.0 micromol/L, 2.0 micromol/L, 10.0 micromol/L). The Ki-67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-thymidine incorporation. The apoptosis of 786-0 cells was detected by TUNEL assay. The control groups were treated with anti-sense oligonucleotide (ASODNs) targeting Ki-67 gene. Our results showed that the Ki-67 expression of 786-0 cells treated with anti-sense PNAs (16.9+/-0.7) was significantly inhibited as compared with that of the control groups (28.6+/-0.4) (P<0.01). The Ki-67 protein rate of 786-0 cells treated with anti-sense PNAs (42.1 +/-2.2) was significantly reduced when compared with that of the control groups (83.6+/- 1.4) (P<0.01). Proliferation of 786-0 cells treated with anti-sense PNAs (20.7+/- 1.5) was significantly inhibited as compared with that of the control groups (58.6+/- 1.4) (P<0.01). The apoptosis rate of 786-0 cells treated with anti-sense PNAs (28.7+/- 2.3) was significantly increased higher compared with that of the control groups (13.8 +/- 1.0) (P<0.01). From these finds we are led to conclude that anti-sense PNAs targeting Ki-67 gene have stronger effects on the inhibition of the proliferation and induction of apoptosis of human renal carcinoma cells than ASODNs targeting Ki-67 gene. The strategies using anti-sense PNAs targeting Ki-67 gene may be a promising approach for the treatment of renal cell carcinoma.
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<p><b>OBJECTIVE</b>To investigate the mechanism of immune escape in renal cell carcinoma(RCC).</p><p><b>METHODS</b>Fas and FasL expressions were examined by immunohistochemical technique in 44 RCC patients, with the Ki67 expression and apoptosis of tumor infiltrating lymphocytes(TIL) monitored simultaneously. Cytokines including IL2 and IFN alpha were used to induce the expression of the renal carcinoma cell lines 786-0 cells. Combination treatment of 786-0 with cytokines and Anti-Fas monoclonal antibody (FasAb) was used to induce apoptosis. FasL function was assessed by in vitro co-culture assays using renal cancer cells 786-0 and Fas-sensitive Jurkat T-cells.</p><p><b>RESULTS</b>(1) Fas expression rate in RCC(22.8%) was lower than that in the controlled normal kidney tissues(53.8%, P < 0.01). FasL expression rate in RCC (46.5%) was higher than that in the controlled normal kidney tissues (23.2%, P < 0.01). That of Ki67 was 32.8%, with the expressions of Fas and Ki67 showing a negative correlation (r = -0.62, P < 0.05). In contrast, the expressions of FasL and Ki67 showed a positive correlation. (r = 0.93, P < 0.01). The Fas expression of stage I was significantly higher than that of stages III and IV. The expression rate of FasL in RCC was significantly increased with RCC stage (P < 0.01). (2) The apoptotic rate of TIL in RCC (33.9%) was significantly higher than that of the normal kidney tissues (3.5%, P < 0.01). The expression of FasL and the apoptotic rate of TIL in RCC gave a positive correlation (r = 0.96, P < 0.01). (3) Fas expression rate in 786-0 cells was 13.7%. The apoptotic rate mediated by FAsAb was 9.6%. IFN alpha was able to up-regulate the Fas expression and subsequently augment the FasAb-mediated apoptosis in 786-0 cells. But IL2 did not show similar effects. (4) The FasL expression rate of 786-0 was 18.6%. FasL expressed by 786-0 cells was able to induce apoptosis of Jurkat T-cells in co-culture assays and the apoptosis of Jurkat T-cells was significantly lowered after blocking the effect of FasL with Fas-neutralizing antibody NOK-2, giving the apoptotic rates of 14.9% and 2.0%, respectively, the difference therein is statistically significant (P < 0.01).</p><p><b>CONCLUSION</b>Down-regulation of Fas expression and up-regulation of FasL-expression are the mechanisms through which the RCC cells escape from immune attack.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Renal Cell , Allergy and Immunology , Fas Ligand Protein , Ki-67 Antigen , Allergy and Immunology , Kidney Neoplasms , Allergy and Immunology , Membrane Glycoproteins , Allergy and Immunology , fas Receptor , Allergy and ImmunologyABSTRACT
Aim To explore the protective effect of Sertoli cells on the co-transplanted allogeneic renal cells in male and female recipient rats. Methods Testicular Sertoli cells were prepared by digestion with trypsin， collagenase and DNase， while renal cells were prepared by digestion with trypsin alone. FasL and Fas expressions were detected respectively by FCM before transplantation. About 106 cells were injected into the allogeneic renal subcapsule. In order to demonstrate the survival of renal cells， the expression of g-PG in graft was examined by SABC method. Apoptosis of the lymphocytes surrounding the graft was observed by TUNEL(terminal deoxynucleotidyl transferase-mediated X-dUTP nick end labeling). Results The grafts were analyzed histologically 20 days after transplantation. The renal cells transplanted alone were all rejected, while the survival rates of the mixed cell transplantation were 87.5％ and 77.8％ in male and female recipients respectively. When the renal cells co-transplanted with Sertoli cells treated by anti-FasL mAb， only 30.0％ grafts were survived. Apoptosis of lymphocytes surrounding the graft were quite evident. Conclusion Sertoli cells expressing FasL can protect renal cells from allograft rejection by inducing apoptosis of Fas expressing T cells surrounding the grafts.
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Objective:To investigate the role of cytokines in modulating expression of Fas and FasL in renal cell carcinoma cells(RCCs) and its implication.Methods:Combination treatment of 786 0 and GRC 1 cells with cytokines including IFN??IFN?,IL 2,TNF? and anti Fas monoclonal antibody (FasAb) to induc apoptosis.FasL function was assessed by coculture assays in vitro using renal cancer cells 786 0 or GRC 1 and Fas sensitive Jurkate T cells.Results:1.Either IFN? or IFN? could up regulate the Fas expression and subsequently augment the Fas mediated apoptosis in 786 0 and GRC 1 cells.2.IFN? and IFN? could up regulate the FasL expression in 780 0 and GRC 1 cells.And subsequently augmente the apoptosis of Jurkat T cells cocultured with 786 0 and GRC 1 cells.IFN? had the same effects on 786 0 cells.IFN? had the same effects on GRC 1 cells.Conclusion:IFN? and IFN? could agument the Fas mediated apoptosis of RCCs by enhancement of Fas expression,but they also up regulate the expression of FasL in RCCs and subsequently augmente the apoptosis of RCCs by enhancment the apoptosis of T lymphocytes by Fas/FasL pathway.
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Objective To investigate the mechanism of renal cell carcinoma (RCC) in affecting the immune system of the host. Methods FasL expression and the apoptosis of tumor infiltrating lymphocytes (TIL) were examined by immunohistochemical technique in 44 cases of RCC.FasL function was assessed by coculture assays in vitro using the renal cancer cells 786 0 or GRC 1 and the Fas sensitive Jurkat T cells. Results (1)FasL expression rate in RCC (46.5%) was higher than that in the normal kidney tissues (23.2%, P