| Objective To investigate the expression of programmed cell death-1(PD-1)m RNA and programmed cell death-ligand1(PD-L1)m RNA in gastrointestinal stromal tumor(GIST)by RNAscope technology and its correlation with several clinicopathological parameters and to compare the difference between PD-1/PD-L1 m RNA and PD-1/PD-L1 protein detected by immunohistochemistry.Methods 50 cases of GIST were collected from the North China University of Science and Technology Affiliated Hospital during 2015 to 2017.8 cases of the normal gastrointestinal mucosa,4 cases of the leiomyoma,4 cases of the schwannoma and 4 cases of the aggressive fibrumatosis were collected as the control group.The expression of PD-1/PD-L1 m RNA in 50 cases of GIST and 20 cases of control group were detected by RNAscope technology.The expression of PD-1/PD-L1 protein in 50 cases of GIST were detected by immunohistochemistry.Results GISTs of 20 men and 30 women with a mean age 57 years(range 45-74 years)were included with tumors arising from 20 cases of the stomach(44.0%)and 28 cases of out of the stomach including with 22 cases of the small bowel(44.0%)?3 cases of the bowel(6.0%)?2 cases of the duodenum(4.0%)and 1cases of the peritoneal cavity(2.0%).In 50 cases of gastrointestinal stromal tumors cases,the expression of PD-1/PD-L1 protein in the positive rate was 44.0%(22/50)and 60.0%(30/50),while the positive rate of PD-1/PD-L1 protein in the control group was 5%(1/20),the difference was statistically significant(P<0.05).In 50 cases of gastrointestinal stromal tumors,PD-1 protein was related with the gender,age and histopathologic type(P>0.05).PD-1 protein is associated with tumor location,tumor size,mitotic counts,the positivity of Ki-67 and the grade of NIH of GIST(P<0.05).The positive rate of patients not in stomach(57.1%,16/28)was higher than that in stomach(27.3%,6/22),statistical analysis the difference was significant(P=0.047);in tumor size group,the positive rate of patients in large size was higher than those in small size and the difference was significant(P=0.001).PD-1 protein has a higher positive expression rate in patients with more mitotic counts and the difference was significant(P=0.001).The positive rate of patients with Ki-67>8%(70.0%,14/20)was higher than that with Ki-67<8%(26.7%,8/30),statistical analysis the difference was significant(P=0.002).In the group of the grade of NIH of GIST,PD-1 protein has a higher positive rate in patients with higher risk and the difference was significant(P=0.001).In 50 cases of gastrointestinal stromal tumors cases,PD-L1 protein was not related to the gender,age and histopathologic type(P>0.05).PD-L1 protein is associated with tumor location,tumor size,mitotic counts,the positivity of Ki-67 and the grade of NIH of GIST(P<0.05).The positive rate of patients not in stomach(92.9%,26/28)was higher than that in stomach(18.2%,4/22),statistical analysis the difference was significant(P=0.001);in tumor size group,the positive rate of patients in large size was higher than those in small size and the difference was significant(P=0.001).PD-L1 protein has a higher positive expression rate in patients with more mitotic counts and the difference was significant(P=0.001).The positive rate of patients with Ki-67>8%(100.0%,20/20)was higher than that with Ki-67<8%(33.3%,10/30),statistical analysis the difference was significant(P=0.001).In the group of the grade of NIH of GIST,PD-1 protein has a higher positive rate in patients with higher risk and the difference was significant(P=0.001).In 50 cases of gastrointestinal stromal tumors cases,the expression of PD-L1/PD-1m RNA,the positive rate was 52.0%(26/50)and 68.0%(34/50),and PD-1/PD-L1 m RNA in the control group were negative,the positive rate was 10.0%(2/20),there was significant difference in statistical significance(P<0.05).In 50 cases of gastrointestinal stromal tumors cases,PD-1m RNA was not related to the gender,age and histopathologic type(P>0.05).It was associated with tumor location,tumor size,mitotic counts,the positivity of Ki-67 and the grade of NIH of GIST(P<0.05).The positive rate of patients not in stomach(78.6%,22/28)was higher than that in stomach(18.2%,4/22),statistical analysis the difference was significant(P=0.001);in tumor size group,the positive rate of patients in large size was higher than those in small size and the difference was significant(P=0.017).PD-1m RNA has a higher positive expression rate in patients with more mitotic counts and the difference was significant(P=0.019).The positive rate of patients with Ki-67>8%(80.0%,16/20)was higher than that with Ki-67<8%(33.3%,10/30),statistical analysis the difference was significant(P=0.001).In the group of the grade of NIH of GIST,PD-1 protein has a higher positive rate in patients with higher risk and the difference was significant(P=0.001).In 50 cases of gastrointestinal stromal tumors cases,PD-L1 m RNA was not related to the gender,age,tumor size and histopathologic type(P>0.05).It was associated with tumor location,mitotic counts,the positivity of Ki-67 and the grade of NIH of GIST(P<0.05).The positive rate of patients not in stomach(85.7%,24/28)was higher than that in stomach(45.5%,10/22),statistical analysis the difference was significant(P=0.002).PD-1m RNA has a higher positive expression rate in patients with more mitotic counts and the difference was significant(P=0.014).The positive rate of patients with Ki-67>8%(90.0%,18/20)was higher than that with Ki-67<8%(53.3%,16/30),statistical analysis the difference was significant(P=0.006).In the group of the grade of NIH of GIST,PD-1 protein has a higher positive rate in patients with higher risk and the difference was significant(P=0.001).In immunohistochemistry and RNAscope detection of PD-1 in 50 cases of gastrointestinal stromal tumors,19 were positive,21 were negative,3 were positive in immunohistochemistry and negative in RNAscope,7 were negative in immunohistochemistry and positive in RNAscope.There was no significant difference between the two methods(P > 0.05).The consistency of the method is general(Kappa value is 0.602).In immunohistochemistry and RNAscope detection of PD-L1 in 50 cases of gastrointestinal stromal tumors,26 were positive,12 were negative,4 were positive in immunohistochemistry and negative in RNAscope,8 were negative in immunohistochemistry and positive in RNAscope.There was no significant difference between the two methods(P > 0.05).The consistency of the method is general(Kappa value is 0.483).Conclusions 1 The positive rates of PD-1/PD-L1 protein and mRNA in patients with gastrointestinal stromal tumors were significantly higher than those in control tissues(including normal gastrointestinal mucosa,leiomyoma,schwannoma and invasive fibroma).2 In patients with gastrointestinal stromal tumors,PD-1 protein and PD-1 m RNA were not related to gender,age and histological classification(P > 0.05),but were related to the location of tumors,size of tumors,mitotic count,Ki-67 index and risk classification(P < 0.05).The positive expression rate of PD-1 protein and m RNA was higher in non-gastric GIST patients;in the grouping of the size of tumors,the positive rate of patients with larger tumors was higher than who with smaller tumors;the positive expression rate of PD-1 protein and m RNA was higher in GIST patients with more mitotic figures;the positive expression rate of PD-1 protein and m RNA was higher in GIST patients with more than 8% Ki-67;in the grouping of risk grading,the positive expression rate of PD-1 protein and m RNA was higher in GIST patients with higher risk grading.3 In patients with gastrointestinal stromal tumors,PD-L1 protein and PD-L1 m RNA were not related to gender,age and histological classification(P > 0.05),but were related to the location of tumors,size of tumors,mitotic count,Ki-67 index and risk classification(P < 0.05).The positive expression rate of PD-L1 protein and m RNA was higher in non-gastric GIST patients;in the grouping of the size of tumors,the positive rate of patients with larger tumors was higher than who with smaller tumors;the positive expression rate of PD-L1 protein and m RNA was higher in GIST patients with more mitotic figures;the positive expression rate of PD-L1 protein and m RNA was higher in GIST patients with more than 8% Ki-67;in the grouping of risk grading,the positive expression rate of PD-L1 protein and m RNA was higher in GIST patients with higher risk grading.4 There was no significant difference between the results of immunohistochemistry and RNAscope in detecting PD-1/PD-L1,and the consistency between the two methods was general.There are limited reports on the use of RNAscope and immunohistochemical methods to detect PD-1 and PD-L1 in GIST cases.More data are needed for comparative analysis.Figure 7,Table 9,Reference 41. |
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