ABSTRACT
@# Objective: To investigate the expression of histone methyltransferase G9a in gastric cancer tissues and its correlation to prognosis, and to observe the effect of G9a inhibitor on the proliferation and apoptosis of gastric cancer cells. Methods: The expression level of G9a in gastric cancer tissues and its correlation to prognosis were analyzed by using the Kaplan-Meier Plotter and Oncomine database. Human gastric cancer cell line SGC-7901 and MKN-45 were selected as study subject. The expression level of G9a protein was detected by Western blotting. The morphological change of gastric cancer cells after the treatment of G9a inhibitor BIX01294 was observed. CCK-8 proliferation experiment and plate colony formation assay were used to examine the proliferation ability and clone formation rate of gastric cancer cells. The changes of cell apoptosis were detected by Annexin-V staining. Results: G9a was highly expressed in gastric cancer tissues (P<0.01), and the high expression of G9a was positively correlated with poor prognosis of gastric cancer patients (P<0.01). After the treatment of BIX01294, the morphology of gastric cancer cells was changed, the volume of gastric cancer cells reduced, the intercellular connections disappeared, and even the apoptotic manifestations appeared, such as the shrinking,, becoming round and cast-off etc. BIX01294 could significantly inhibit the proliferation and colony formation but promote the apoptosis of gastric cancer cells (all P<0.05). Conclusion: Histone methyltransferase G9a was highly expressed in gastric cancer tissues, and its high expression level was positively correlated with poor prognosis. The proliferation of gastric cancer cells was obviously inhibited while the apoptosis was significantly promoted after inhibiting G9a expression.
ABSTRACT
Objective:To establish a mouse model of gastric cancer by inoculating MKN45 cells into mice with normal immune function utilizing microcarrier technology. Methods:A total of 60 male C57BL/6 mice were randomly divided into three groups, namely, 2D, con-trol, and 3D groups, according to the coculture system of MKN45 and microcarrier. The mouse models of gastric carcinoma were estab-lished by hypodermic injection. The time of tumorigenesis, rate of tumor formation, and pathological features were observed in each group. Results:In the 3D group, the time of tumor formation was short, whereas the rate of tumor formation was high (80%). No de-tectable tumor formations were observed in the 2D and control groups. HE and immunohistochemical staining of the transplantation tumor model showed evident characteristics of human gastric cancer. Conclusion:A human gastric cancer model in normal immune mice was successfully established. The onset and development mechanism of gastric cancer could be more effectively investigated in mice with normal immune function through this model. Moreover, a more valuable and new animal model for the research and devel-opment of anticancer drug was established.
ABSTRACT
Objective To study the effect of annonaceous acetogenins (ACGs) on human gastric cancer cells in vitro. Methods After ACGs were administered to gastric cancer cells in vitro, the cell viability, cell adhesion ability and cell migration ability were assessed by MTT assay, adhesion assay and wound-healing assay, respectively. Results ACGs inhibited the cell viability, adhesion ability and migration ability in a dose-dependent manner in gastric cancer cells. Conclusion ACGs could inhibit cell activities of human gastric cancer cells in viro, and will be developed as a promising anticancer candidate and used in gastric cancer.
ABSTRACT
Objective To investigate the relevant mechanism of different concentrations of Shenqi YiliuDecoction medicated serum on blocking cell cycle and inhibiting invasion and metastasis of gastric cancer cell lines MKN-45. Methods Sixty SPF Wistar rats were randomized intoShenqi Yiliu Decoction low-, medium-, and high-dose groups and control group, with 15 rats in each group. Low-, medium- and high-dose groups were fed with different concentrations of TCM liquid from which containing crude medicine (0.25, 0.50, 1.00 g/mL) separately for gavage. Rats in the blank group were given the same amount of drinking water for gavage, twice a day, for 7 days.2 h after the last gavage, rat blood was taken and serum was separated. After MKN-45 cells were dealt with different concentrations of medicinal serum, FCM was used to detect cell cycle and immunohistochemistry technology was used to detect the expressions of COX-2 and PTEM proteins.Results FCM analysis showed that medicated serum could increase G0-G1 phase and shorten S phase of cells; medicated serum could reduce the positive expression rate of COX-2 of the MKN-45 cells and increase positive expression rate of PTEN proteins (P<0.05).ConclusionShenqi YiliuDecoction medicated serum can block the cell cycle and inhibit invasion and metastasis of MKN-45 cell lines, which may be related to the intervention in the expression levels of COX-2 and PTEN proteins.
ABSTRACT
Aim To establish the adherence of H pylori to MKN45 cell model for screening and evaluating anti-adhesive drugs.Methods H pylori suspension was incubated with FITC solution in different conditions(FITC concentration and incubation time)to optimize the labeling conditions.At the same time,Gram staining was used to determine the optimal adhesion time of H pylori to MKN45 cells.Results H pylori suspension(1?106 cuf?ml-1)incubating with FITC(5 mg?L-1)for 1 h could achieve the optimal effect for FITC-labeling H pylori.Besides,Gram staining indicated that the optimal adhesion time for H pylori to MKN45 cells was 1 h.Based on the model,the crude Aloe polysaccharide was evaluated for its anti-adhesive activity and the results showed it could significantly reduce the adherence of H pylori to MKN45 cells at the concentration of 1 g?L-1(P