CDKN1A, 1B, TP53 mRNA and miR-34a, 192, 215, 221 Expression of Crypts and Stroma in Human Colorectal Cancer
Hiroyuki Nozaka, Akifumi Mayama, Tomisato Miura, Hideki Takami, Koudai Takahashi, Yasuhiro Konishi, Yoshiharu Mue, Masamichi Suzuki, Noriyuki Uesugi, Kazuyuki Ishida, Tamotsu Sugai. Hirosaki University, Hirosaki-shi, Aomori, Japan; Iwate Medical University, Morioka-shi, Iwate, Japan
Background: The carcinogenesis of colon is caused by accumulation of genetic alternation or mutation. Progress of human cancers is relevant to activation or inactivation of both oncogenes and cancer suppressor genes. Resent works have shown significant genomic DNA biomarkers, it is also reported the posttranscriptional regulation of gene by mRNAs or non-coding RNA. It is suggested that the functional mRNAs and miRNAs status are significance for clarification of cancer progress. However, it is not reported that mRNA/miRNA expression status as the clinicopathological biomarker in only crypts or stroma. In this study, we examined CDKN1A, 1B, TP53 mRNA and miR-34a, 192, 215, 221 expression of crypts/stroma in human colorectal cancer, and analized significance as the clinicopathological biomarker.
Design: Tumors were collected from 30 patients diagnosed with primary advanced colorectal cancer. After the surgery extraction, we collected a part of both normal and tumor tissues from fresh colon or rectum. Tissues were sliced with a razor into minute pieces, half of pieces were dissolved in TRIzol reagent (Whole). Crypt was isolated from the left-behind pieces by crypt isolation method, and dissolved in TRIzol reagent (Crypt). After extraction of total RNA, and CDKN1A, 1B, TP53 mRNA and miR-34a, 192, 215, 221 expression in crypts/whole were measured by qRT-PCR. The relative quantification values compared with normal tissues were calculated, and statistical analysis was carried out between pTMN StageI+II and pTMN Stage III+IV group by SPSS.
Results: CDKN1A, miR-192, 215 showed down regulation in both crypt and whole, and miR-34a showed down regulation in only whole. Statistical analysis was shown in table1. In the analysis of crypt, CDKN1A showed significantly down regulation in Stage III+IV group. In the analysis of whole, mir-34a and 221 showed significantly down regulation in Stage III+IV group.