| Objective: We researched the relationship between the differential proteomic expression of RORαprotein in diallyl disulfide-induced human gastric cancer cells and human gastric cancer cells differentiation base on previous study. We hope to supply a theoretical evidence for searching a new target of gastric carcinoma. Results: 1.As exposed to inversion microscope and optics microscope, when DADS was added to culture medium 24h after cell seeding was found to severely reduce the number of adherent cells in the culture flask of cell culture when compared with cells cultured in the presence of vehicle alone. Hematoxylin and eosin coloration showed that human gastric cancer cells that still adhere the flask showed malignance declined as nuclear-to-cytoplasmic ratio was reliable, the number of nucleoli in the cell nuclear decreased, size of nucleus shortened, and coloring of nucleus turned weak.2.After MGC803, BGC823, SGC7901 and MKN28 cells treated with DADS for 24 hours, Immunocytochemical stain indicated that positive expression of RORαincreased compared with the control.3.By means of PI staining flow cytometry the effect of RORαin MGC803, BGC823, SGC7901 and MKN28 cells treated with DADS 24h were observed. The expression rate of RORαin four tumor cells were 9.0±1.20, 7.4±1.32, 26.0±0.14 and 5.3±1.32, compare with 6.0±1.20, 4.4±0.8, 12.4±0.02 and 5.0±0.29 as a control. The expression rate of the treated group were elevated than the control group, especially in SGC7901 and MGC803 (P <0.05). The fluorescence intensity showed that four human gastric cancer cells treated with DADS for 24h, were 35.11±6.34, 26.14±3.40, 37.65±6.08 and 29.44±4.29, respectively, compare with 13.90±1.80, 9.6±2.88, 15.32±4.08 and 13.45±3.81 as a control (P <0.01 or P <0.05). The fluorescence intensity of the treated group was elevated than the control group.4.After treated with DADS for 24h, RT-PCR analysis showed that the RORαexpression were up-regulated, the gray scale value ratio of RORα/β-actin were 1.09±0.01 and 0.97±0.01 in SGC7901 and MGC803, compare with 0.45±0.03 and 0.44±0.02 as a control (P <0.01), 1.01±0.02 and 0.91±0.01 in BGC823 and MKN28, compare with 0.68±0.01 and 0.72±0.01 as a control (P <0.05).5.After MGC803, BGC823, SGC7901 and MKN28 cells treated with DADS for 24h, Western Blot showed that the RORαexpression was increased, the gray scale value ratio of RORα/β-actin were 0.71±0.03, 0.69±0.01, 0.80±0.02 and 0.79±0.01; 0.87±0.01 ,0.86±0.01, 0.93±0.01 and 0.88±0.02; 1.31±0.01, 1.14±0.04, 1.44±0.03 and 0.96±0.02 respectively, compare with 0.32±0.01, 0.43±0.01, 0.38±0.01 and 0.68±0.02 as a control (P <0.05).Conclusion: 1.DADS can up-regulate RORαprotein and induce MGC803, BGC823, SGC7901 and MKN28 cells differentiation.2.DADS can induce human gastric cancer MGC803, BGC823, SGC7901 and MKN28 cells differentiation through up-regulate RORαprotein. Objective: To make clear the RORαprotein play in occurrence, development of gastric carcinoma, we intended to detect the expression of them in gastric carcinoma tissue chip. We hope to supply experimental evidence for searching a tumor marker of gastric carcinoma.Methods: We applied tissue chip technique and SP Immunohistochemistry method to detect RORαprotein expression in the 90 cases gastric carcinoma, the 48 cases tissue nearby and 22 cases matched non-cancer mucosa and evaluate the influence of abnormal expression of RORαprotein for incidence and progression of gastric carcinoma.Results: The result of facture TMA: The 90 cases of gastric carcinoma, 48 cases of tissue nearby and 22 cases matched non-cancer mucosa of our recipient block was exquisite and had no splits. The chip cores were arraged orderly and clearly in the block. The chip cores had no shift and distort and obviously rugate in slides when they cut from block. They were answered for the request of research.Immunohistochemistry results revealed that: from the 90 cases, the positive rates of RORαprotein expression in matched non-cancer mucosa, tissue nearby and gastric carcinoma with 83.36%(19/22), 56.25%(27/48)and 24.44%(22/90), respectively. The RORαprotein positive expression in gastric carcinoma were significantly lower than those matched non-cancer mucosa and the tissue nearby(P <0.05).The positive rate of RORαprotein expression were 45.00%(9/20)in well differentiated, 27.59%(8/29)in moderate differentiated, 14.29%(3/21)in poor differentiated gastric carcinoma, 9.05% ( 1/11 ) in mucinous adenocarcinoma and11.11%(1/9)in signet ring cell carcinoma respectively, and the positive rates of RORαprotein expression in well differentiated were significantly higher than poor differentiated gastric carcinoma(P <0.05).Conclusion: 1.The positive rate of RORαprotein expression in gastric carcinoma was significantly lower than in normal gastric tissue and tissue nearby, which indicated the down-regulation of RORαprotein was related to the incidence and development of gastric carcinoma.2.The positive rate of RORαprotein expression in well differentiated were significantly higher than poor differentiated gastric carcinoma, which indicated RORαprotein maybe related to the degree of differentiation of gastric carcinoma. |
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