| Objective:By detecting the expression of IL-17 D gene in epithelial ovarian cancer, cervical cancer, endometrial carcinoma, gastric cancer tissues and their own controls tissues and the expression of the membranous form(be MICA, m MICA) of MHC class I-related protein A(MICA) on the cell surface of these tissues respectively and measuring the expression of the soluble form(soluble MICA, s MICA) of MHC class I-related protein A(MICA) in peripheral blood serum of epithelial tumor patients and the healthy people,we preliminary to discuss the the effect of IL-17 D gene in the NKG2D- MICA immune pathways mediated by NK cells.Methods:1 Charge epithelial ovarian cancer tissue specimens of 36 cases, 50 cases of cervical cancer tissue samples, endometrial tissue specimens of 48 cases, 28 cases of gastric epithelial tissue specimens and 10 cases of specimens of their respective control group. These specimens were confirmed by pathology and charged from patients of the fourth hospital of hebei medical university from October 2013 to October 2014.The tissues of the control group(normal cervical tissue, endometrial tissue and ovarian tissue)were removed from the patients with uterine fibroid, and the control stomach tissues were apart from the margins of gastric cancer stomach tissue of 5 cm. Double each sample specimen, every specimen used for q RT-PCR and Flow cytometry detection were to be stored in liquid nitrogen within 0.5 h and- 80 preservation℃.2 Charge the serum of patients of four epithelial tumors and 10 cases of physical health adult by contrast, and all the members were dawn fasting serum. These serum were charged from peripheral blood 2m L with anticoagulant tubes, and were centrifugaled within 2h,- 80 ℃preservation for ELISA method detection.3 IL-17 D m RNA in four types of epithelial tumors in the organization and the control group was determined by q RT-PCR analysis, The serum levels of s MICA and m MICA expressed on tumor cells was assessed by ELISA and Flow cytometry analysis. Use SPSS 13.0 statistical analysis software to analyze experimental data. P<0.05 indicates that the difference had a statistical significance.Results: 1 The results of q RT- PCR showed that expression of IL-17 D gene in ovarian epithelial tissues was about 32 times of the normal ovarian tissue and 7 times, 7 times, 64 times of the other control tissues respectively. All the differences had a statistical significance(P<0.05). 2 Flow cytometry analysis suggested that the expression of MICA on the surface of epithelial ovarian cancer was 223.68±3.06, which was obviously higher than the control group 11.63±1.95, the expression of MICA on the surface of cervical cancer was 258.97±3.40, which was obviously higher than the control group 10.26±1.67, the expression of MICA on the surface of endometrial ovarian cancer was 278.40±3.43, which was obviously higher than the control group 13.41±1.32, the expression of MICA on the surface of epithelial gastric cancer was 347.76±3.46, which was obviously higher than the control group 69.42±1.87. The differences were statistically significant(P<0.05). 3 The serum levels of s MICA(ng/m L) measured by ELISA in the group of epithelial ovarian cancer were 52.13±2.63 and the control group 39.26±1.37, which suggested the level of s MICA in epithelial ovarian cancer group was higher than in the control group(P<0.05), The serum levels of s MICA were no obvious difference in the group of cervical cancer 40.17±3.19 and the control group 39.26±1.37(P>0.05), The serum levels of s MICA were no obvious difference in the group of endometrial cancer 40.60±2.65 and the control group 39.26±1.37(P>0.05), The serum levels of s MICA in the group of epithelial gastric cancer were 63.28±2.44 and the control group 39.26±1.37 respectively, which suggested the level of s MICA in epithelial gastric cancer group was higher than in the control group(P<0.05). 4 The serum level of s MICA(?X±s ng/m L)、the expression of m MICA(?X ± s) and IL-17D( 2-ΔΔCт) in m RNA in epithelial ovarian cancer 4.1 There were 19 cases of positive Lymph node metastasis and 17 cases of negative Lymph node metastasis in 36 cases epithelial ovarian cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the positive and negative group were statistically significant(P<0.05). 4.2 There were 15 cases of stage Ⅰ~ Ⅱand 21 cases of stage Ⅲ~ Ⅳrespectively in the group of epithelial ovarian cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the two groups were statistically significant(P<0.05). 4.3 There were 25 cases of the poorly differentiaed patients and 11 cases of the higher/moderately differentiated patients respectively in epithelial ovarian cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the two groups were statistically significant(P<0.05). 4.4 There were 9 cases of endometrioid cancer, 8 cases of serous cancer, 13 cases of poorly differentiated carcinoma, 6 cases of mucinous cancer respectively. Compared with each other, there were no obvious difference of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA in different histological type of epithelial ovarian cancer(P>0.05). 5 The serum level of s MICA(?X±s ng/m L)、the expression of m MICA(?X ± s) and IL-17D( 2-ΔΔCт) in m RNA in cervical cancer 5.1 There were 30 cases of stage and 20 cases of stage respectively in Ⅰ Ⅱ50 cases of cervical cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the two groups were not statistically significant(P>0.05). 5.2 There were 29 cases of the poorly differentiaed patients and 21 cases of the higher/moderately differentiated patients respectively in cervical cancer. The differences of the expression of m MICA and IL-17 D in m RNA between the two groups were statistically significant(P<0.05). But the differences of serum level of s MICA were not statistically significant(P>0.05). 5.3 There were 32 cases of squamous-cell cancer, 10 cases of adenocarcinoma and 8 cases of other types cancer. Compared with each other, there were no obvious difference of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA in different histological type of cervical cancer(P>0.05). 6 The serum level of s MICA(?X±s ng/m L)、the expression of m MICA(?X ± s) and IL-17D( 2-ΔΔCт) in m RNA in endometrial cancer 6.1 There were 45 cases of stage Ⅰ~ Ⅱand 3 cases of stage Ⅲ~ Ⅳrespectively in 48 cases of endometrial cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the two groups were not statistically significant(P>0.05). 6.2 There were 27 cases of the poorly differentiaed patients and 21 cases of the higher/moderately differentiated patients respectively in endometrial cancer. The differences of the expression of m MICA and IL-17 D in m RNA between the two groups were statistically significant(P<0.05). But the differences of serum level of s MICA were not statistically significant(P>0.05). 6.3 There were 40 cases of adenocarcinoma, 8 cases of other types cancer respectively. There were no obvious difference of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA in different histological type of endometrial cancer(P>0.05). 7 The serum level of s MICA(?X±s ng/m L)、the expression of m MICA(?X ± s) and IL-17D( 2-ΔΔCт) in m RNA in epithelial gastric cancer 7.1 There were 12 cases of positive Lymph node metastasis and 16 cases of negative Lymph node metastasis in 28 cases epithelial gastric cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the positive and negative group were statistically significant(P<0.05). 7.2 There were 8 cases of stage Ⅰ~ Ⅱand 20 cases of stage Ⅲ~ Ⅳrespectively in the group of epithelial gastric cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the two groups were statistically significant(P<0.05). 7.3 There were 15 cases of the poorly differentiaed patients and 13 cases of the higher/moderately differentiated patients respectively in epithelial gastric cancer. The differences of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA between the two groups were statistically significant(P<0.05). 7.4 There were 4 cases of squamous-cell cancer and 24 cases of adenocarcinoma. There were no obvious difference of serum level of s MICA, the expression of m MICA and IL-17 D in m RNA in different histological type of epithelial gastric cancer(P>0.05).Conclusion: IL-17 D can possiblely promote tumor growth in the four epithelial tumors. The mechanism may be that IL-17 D can down-regulated m MICA on the surface of cancer cells, and enhanced the expression of s MICA, so as leads to the immune kill pathway of NKG2D-MICA mediated by NK cell to cancer cells decreases in epithelial ovarian cancer and gastric cancer. In this way, tumor can escape from immune surveillance, and acquire tumor immune escape. The experiment can not prompt this mechanism also involves in the tumor immune escape in cervical cancer and endometrial cancer. As for the complex mechanism, it is worth our further discussion. |
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