Tumor Weight In Control Group Research Articles

Fat1 inhibits cell proliferation via ERK signaling pathway in esophageal squamous cell carcinoma

Objective: To clarify the mechanism of Fat1 on the proliferation of esophageal squamous cell carcinoma (ESCC). Methods: KYSE450 cells were transfected with Plko.1-puro-GFP-shRNA-Fat1 plasmid and real time polymerase chain reaction (PCR) was used to verify the efficiency of Fat1 knockdown. The effects of Fat1 and extracellular regulated protein kinase (ERK) pathway inhibitor U0126 on the proliferation of ESCC cells were detected by methyl thiazolyl tetrazolium (MTT). Colony formation assay was used to detect the colony formation ability. Cell cycle was detected by live cell imaging. Western blot was used to observe the level of target protein. Mouse xenograft assay was applied to detect the effect of Fat1 knockdown on KYSE450 cell tumor growth. Immunohistochemistry was used to detect the expressions of related proteins in tumor sections. Results: The efficiency of Fat1 knockdown was (77.1±6.9)% in Fat1 sh1 group and (77.7±7.1)% in Fat1sh2 group. Compared with the control group, the cell proliferation and the expression of p-ERK1/2 were significantly increased in Fat1 sh1 and Fat1sh2 group (P<0.05). After U0126 treatment, the effect of Fat1 knockdown on the proliferation of KYSE450 cells disappeared, and the expression of p-ERK1/2 in KYSE450 cells decreased to a level similar to that in the control group. The number of cell clones in the control group was (72±8), lower than (155±28) and (193±9) in the Fat1sh1 and Fat1sh2 groups, respectively (P<0.05). In KYSE450 cell, division time was shortened from 1 622±32 min in control group to 1 408±29 min in Fat1 sh1 group, the difference was statistically significant (P<0.05). Compared with the control group, the tumor volume of Fat1 knockdown group increased significantly. The tumor weight of control group and Fat1 knockdown group were (0.224±0.028) g and (1.532±0.196) g, respectively, at 4 weeks after inoculation, and the difference was statistically significant (P<0.05). Conclusion: Fat1 inhibits cell proliferation via ERK signaling in ESCC.

Abstract 3751: The potential role of CXCR4 in cisplatin associated resistance in esophageal carcinoma

Abstract Introduction CXC chemokine receptor 4 was found dys-regulated in many different types of human cancers and correlated with poor prognosis of tumor metastasis and resistance to chemotherapy. Limited data show the effect of CXCR4 on the esophageal carcinoma cells, especially with influence of chemotheraputuc reagent-cisplatin. Methods CXCR4 was detected by FACs analysis in esophageal cell lines OE19, OE21, OE33, PT1590 and LN1590. Corronsponding cisplatin resistant cells were established after selection of cisplatin treatment and further confirmed by IC50 and ERCC1. SDF-1α and AMD3100 were used as CXCR4 stimulattion or inhibition. Subcutanous model of OE19 injection in Balb/c nude mice was established and followed by treatment with cisplatin twice a week via i.p injection. Tumors were collected for the further evaluation. Results Esophageal cell line OE19, PT1590 and LN1590 have detetable amount of CXCR4+ subpopulation cells as high as 2.8%±0.9, 0.7%± 0.9 and 1.0%±0.6, respectively. OE19, PT1590 and LN1590 cisplatin resistant subline displayed increased IC50 value of 48h cisplatin treatment from 6.0±1.3 to 12.2±1.0, 1.4±0.7 to 9.1±0.7 and 1.4±0.6 to 7.2±0.6 ug/ml along with a significant overexpression of ERCC1. Interestingly only OE19 and LN1590 cisplatin resistant cell lines showed a significant enreichment of CXCR4+ cells ( 12.1%± 2.5 Vs 2.8%±0.9; 4.2%± 1.1 Vs1.0%±0.6, p&amp;lt;0.05) as compared to parental cells. Meanwhile, side population cells were also increased in those cisplatin resistant variants. There is no difference on tumor weight in control group and cisplatin treatment group in vivo. The relative expression of CXCR4 will be further evaluated by realtime PCR on mRNA level and IHC on protein level. Conclusion Overexpression of CXCR4 is significantly associated with cisplatin-based chemotherapy resistance and might be a prognostic factor in esophageal cancer. Inhibition of CXCR4 might be a strategy to address the chemoresistance of EAC cell lines. Citation Format: Yue Zhao, Yan Wang, Christiane Bruns. The potential role of CXCR4 in cisplatin associated resistance in esophageal carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3751. doi:10.1158/1538-7445.AM2014-3751

Alpha fetal protein-specific hepatocellular carcinoma vaccines inhibit hepatocellular carcinoma growth in nude mice bearing human hepatocellular carcinoma

Objective To study the posibility of hepatocellular carcinoma HCC) vaccines from dendritic cells (DCs) stimulated with alpha fetal protein (AFP) peptides or Lenti-AFP for hepatoceHular carcinoma (HCC) immunotherapy.Methods We created a lentivirus expressing AFP (Lenti-AFP) and AFP peptides liver tumor vaccines,and antitumor activity of AFP-specific HCC vaccines for tumor growth and related cytokines [interleukin (IL)-2,interferon (IFN)-γ,IL-10,tumor necrosis factor (TNF)-αt,perforin and granzyme B,FasL) were observed in 4 groups (Lenti-AFP,AFP1,AFP542 and control) in vivo.Results The AFP-specific tumor vaccines could efficiently kill HCC cells in vivo.The tumor diameter in control group was (12.28 ±3.49) cm,and that in treatment groups was (7.03 ±2.78),(11.00 ±2.57),(8.87 ±3.49) cm,respectively (FD =6.13,P＜0.01).The tumor weight in control group was (0.478 ± 0.225) g,and that in treatment groups was (0.398 ± 0.125),(0.298 ± 0.099),(0.228 ±0.132) g respectively (Fw =5.38,P ＜ 0.01).The levels of IL-2,IFN-γ,TNF-α,perforin and granzyme B were increased to varying degrees,and those of IL-10 were significantly reduced in the treatment groups (P ＜0.05).Conclusion AFP-specific CD8 +/CD4 + T cells tumor vaccines which were activated by either Lenti-AFP-engineered or AFP peptide-pulsed DCs have obviously antitumor activity in vivo.This study provides new insight into the design of HCC immunotherapy. Key words: Carcinoma,hepatocellular; Alpha-fetal protein; Dendritic cell

Depletion of telomerase RNA inhibits growth of gastrointestinal tumors transplanted in mice

To explore effects of telomerase RNA-targeting phosphorothioate antisense oligodeoxynucleotides (PS-ASODN) on growth of human gastrointestinal stromal tumors transplanted in mice. A SCID mouse model for transplantation of human gastrointestinal stromal tumors (GISTs) was established using tumor cells from a patient who was diagnosed with GIST and consequently had been treated with imatinib. GIST cells cultured for 10 passages were used for inoculation into mice. Transfection of PS-ASODN was carried out with Lipotap Liposomal Transfection Reagent. GISTs that subsequently developed in SCID mice were subjected to intra-tumoral injection once daily from day 7 to day 28 post-inoculation, and mice were divided into the following four groups according to treatment: PS-ASODN group (5.00 μmoL/L of oligonucleotide, each mouse received 0.2 mL once daily); imatinib group (0.1 mg/g body weight); liposome negative control group (0.01 mL/g); and saline group (0.01 mL/g). On day 28, the mice were sacrificed, and tumor attributes including weight and longest and shortest diameters were measured. Tumor growth was compared between treatment groups, and telomerase activity was measured by enzyme-linked immunosorbent assay. Apoptosis was examined by flow cytometry. Real-time polymerase chain reaction was used to detect expression of the mRNA encoding the apoptosis inhibition B-cell leukemia/lymphoma 2 (bcl-2) gene. In the PS-ASODN group, tumor growth was inhibited by 59.437%, which was markedly higher than in the imatinib group (11.071%) and liposome negative control group (2.759%) [tumor inhibition = (mean tumor weight of control group--mean tumor weight of treatment group)/(mean tumor weight of control group) × 100%]. Telomerase activity was significantly lower (P < 0.01) in the PS-ASODN group (0.689 ± 0.158) compared with the imatinib group (1.838 ± 0.241), liposome negative control group (2.013 ± 0.273), and saline group (2.004 ± 0.163). Flow cytometry revealed that the apoptosis rate of tumor cells treated with PS-ASODN was 20.751% ± 0.789%, which was higher (P < 0.01) than that of the imatinib group (1.163% ± 0.469%), liposome negative control group (1.212% ± 0.310%), and saline group (1.172% ± 0.403%). Expression of bcl-2 mRNA in the transplanted GISTs was markedly downregulated (P < 0.01) in the PS-ASODN group (7.245 ± 0.739) compared with the imatinib group (14.153 ± 1.618) and liposome negative control group (16.396 ± 1.351). PS-ASODN can repress GIST growth, mediated perhaps by inhibition of telomerase activity and downregulation of bcl-2 expression.

In vivo antitumor activity of hexamethylmelamine against human breast, stomach and colon carcinoma xenografts.

We have evaluated the antitumor activity of Altretamine (hexamethylmelamine, HMM) on human carcinoma xenografts serially transplanted in nude mice. Five human breast carcinoma xenografts, MX‐1, T‐61, MCF‐7, R‐27 and Br‐10, were inoculated subcutaneously into female nude mice. Two human stomach carcinoma xenografts, SC‐1‐NU and St‐4, and three human colon carcinoma xenografts, Co‐3, Co‐4 and Co‐6, were inoculated subcutaneously into male nude mice. One pellet of 17β‐estradiol (0.1 mg/mouse) was inoculated subcutaneously in the mice transplanted with MCF‐7 when the tumors were inoculated. HMM was administered per os daily for 4 weeks. MX‐1 and T‐61 tumors regressed completely after treatment with HMM at a dose of 75 mg/kg (the maximum tolerated dose: MTD) for MX‐1 and 25 mg/kg for T‐61. Br‐10 was sensitive, whereas MCF‐7 and R‐27 were resistant to HMM at its MTD. HMM exerted the most potent antitumor effect against T‐61. Against MX‐1, it exerted an antitumor effect equivalent to that of cisplatin or cyclophosphamide. In addition, this agent was effective against all stomach and colon carcinoma xenografts, in particular St‐4 (T/C%= 10.7: the mean tumor weight of treated group/the mean tumor weight of control group) and Co‐3 (T/C%=31.5%) which are insensitive to presently available agents. HMM seems worthy of further clinical investigation as a candidate agent to treat breast, stomach, colon and other carcinomas.