Identification and characterization of miR-31 potential targets by RNA-seq.

In our previous work, miR-31 displayed differential significant expression in Apostichopus japonicus sea cucumber with skin ulcer syndrome and modulated coelomocytes ROS production by targeting p105. To identify other promising targets ofmiR-31, 4 transcriptome libraries of coelomocytes, as well as 2 control libraries, were constructed frommiR-31 mimics (31 M) or AMO-miR-31 (31I) and injected into a sea cucumber at 12 and 24 h. A total of207,977 unigenes with an average length of 363 bp were assembled, in which17,204 distinct sequences (8.27% of the unigenes) were successfully matched with annotated protein sequences. Fragments per kilobase of transcript per million fragments mapped analysis indicated that 1325 unigenes displayed up-regulated expression profiles in the 31I-12 group and were depressed in the 31M - 12 group compared with the control group. A total of 1470 unigenes showed down-regulated expressions in 31I-12 and were induced in 31 M-12. Similarly, 2079 and 2098 unigenes were detected at 24 h post-injection. Among these unigenes, 36 unigenes (depressed expression in the 31 M group and induced in the 31I group) showed consistent expression patterns at 2 examined time points and were considered promising targets of miR-31. qPCR analysis confirmed that all 4 unigenes showed opposite expression profiles to miR-31 in cultured coelomocytes. Our present work provided a fast and feasible method of identifying miR-31 targets by transcriptome analysis. The results of this study would enhance our present understanding ofmiR-31 function insea cucumber immune regulation.

Expression of Survivin in colorectal cancer tissue and its relationship with clinicopathological features / 中国医师进修杂志

Objective To explore the expression of Survivin in colorectal cancer tissue and its relationship with clinicopathological features.Methods In situ hybridization was used to examine the expression of Survivin mRNA,and immunohistochemical method was used to detect the expression of Survivin protein in 60 cases of colorectal cancer and 30 cases of colorectal normal mucos tissue.The relationship of the expression of Survivin protein with clinicopathological features of colorectal cancer was analyzed.Results The positive expression rate of Survivin protein in colorectal normal mucos tissue (10.00％,3/30) was lower than that in colorectal cancer tissue (73.33％,44/60),and there was significant difference (P ＜ 0.05).The positive expression rate of Survivin mRNA in colorectal normal mucos tissue (6.67％,2/30) was lower than that in colorectal cancer tissue (63.33％,38/60),and there was significant difference (P ＜ 0.05).The expression of Survivin mRNA was positively correlated with depth of invasion,lymph node metastasis,liver metastasis,peritoneal micrometastasis and TNM stage in patients with colorectal cancer (P ＜ 0.01),but had no correlation with gender,age,tumor size,location,degree of differentiation and histological type (P ＞ 0.05).Conclusions The specific high expression of Survivin protein has a high correlation with the occurrence,development,infiltration of colorectal cancer.Survivin may be used as a marker for prognosis of patients with colorectal cancer.