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. 2012 Mar;3(3):560-566.
doi: 10.3892/etm.2011.436. Epub 2011 Dec 28.

microRNA-192, -194 and -215 are frequently downregulated in colorectal cancer

Yeunpo Chiang  1 , Yongxi Song, Zhenning Wang, Zhuangkai Liu, Peng Gao, Jiwang Liang, Jinliang Zhu, Chengzhong Xing, Huimian Xu
Affiliations

microRNA-192, -194 and -215 are frequently downregulated in colorectal cancer

Yeunpo Chiang et al. Exp Ther Med. 2012 Mar.

Abstract

microRNAs (miRNAs) are small, non-coding RNAs of endogenous origin. They have been increasingly shown to have aberrant expression in a number of tumor types. miR-192, -194 and -215 have not been comprehensively investigated using a large number of cases in colorectal cancer (CRC). We extracted total RNA from 107 CRC tissues and three CRC cell lines. Following polyadenylation and reverse transcription, the expression levels of miR-192, -194 and -215 were determined for evaluation of the association between expression levels and clinicopathological characteristics by a quantitative real-time polymerase chain reaction (real-time PCR) method. Finally, we studied the impact of miR-194 on cell proliferation in HCT-116 cells by MTT assay. miR-192, -194 and -215 were significantly downregulated in CRC tissues (all p<0.001, paired t-test) and cancer cell lines (all p<0.05) compared to non-tumor counterparts. Moreover, the expression levels of miR-192, -194 and -215 were demonstrated to be associated with increased tumor sizes (p=0.027, p=0.018, and p=0.027, respectively; Mann-Whitney U test). Also, there were marked correlations among these miRNAs in CRC tissues (all p<0.001, Pearson's regression analysis). Furthermore, we found that the overexpression of miR-194 could significantly inhibit cell proliferation in HTC-116 cells. miR-192, -194 and -215 may be important biological markers as tumor suppressors in the carcinogenesis of CRC.

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Figures

Figure 1.
Figure 1.
Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented as log2 of fold-change of CRC tissues relative to matched NATs. (B, D and F) miR-192, -194 and -215 were differentially expressed between CRC tissues and NATs. These miRNAs were normalized by U6RNA. ΔCt=Ct miRNAs−Ct U6RNA. The ΔCt of these miRNAs was significantly higher in CRC tissues than NATs (p<0.001, paired t-test). NATS, non-tumor adjacent tissues.
Figure 1.
Figure 1.
Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented as log2 of fold-change of CRC tissues relative to matched NATs. (B, D and F) miR-192, -194 and -215 were differentially expressed between CRC tissues and NATs. These miRNAs were normalized by U6RNA. ΔCt=Ct miRNAs−Ct U6RNA. The ΔCt of these miRNAs was significantly higher in CRC tissues than NATs (p<0.001, paired t-test). NATS, non-tumor adjacent tissues.
Figure 1.
Figure 1.
Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented as log2 of fold-change of CRC tissues relative to matched NATs. (B, D and F) miR-192, -194 and -215 were differentially expressed between CRC tissues and NATs. These miRNAs were normalized by U6RNA. ΔCt=Ct miRNAs−Ct U6RNA. The ΔCt of these miRNAs was significantly higher in CRC tissues than NATs (p<0.001, paired t-test). NATS, non-tumor adjacent tissues.
Figure 1.
Figure 1.
Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented as log2 of fold-change of CRC tissues relative to matched NATs. (B, D and F) miR-192, -194 and -215 were differentially expressed between CRC tissues and NATs. These miRNAs were normalized by U6RNA. ΔCt=Ct miRNAs−Ct U6RNA. The ΔCt of these miRNAs was significantly higher in CRC tissues than NATs (p<0.001, paired t-test). NATS, non-tumor adjacent tissues.
Figure 1.
Figure 1.
Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented as log2 of fold-change of CRC tissues relative to matched NATs. (B, D and F) miR-192, -194 and -215 were differentially expressed between CRC tissues and NATs. These miRNAs were normalized by U6RNA. ΔCt=Ct miRNAs−Ct U6RNA. The ΔCt of these miRNAs was significantly higher in CRC tissues than NATs (p<0.001, paired t-test). NATS, non-tumor adjacent tissues.
Figure 1.
Figure 1.
Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented as log2 of fold-change of CRC tissues relative to matched NATs. (B, D and F) miR-192, -194 and -215 were differentially expressed between CRC tissues and NATs. These miRNAs were normalized by U6RNA. ΔCt=Ct miRNAs−Ct U6RNA. The ΔCt of these miRNAs was significantly higher in CRC tissues than NATs (p<0.001, paired t-test). NATS, non-tumor adjacent tissues.
Figure 2.
Figure 2.
Expression levels of miR-192, -194 and -215 in three colorectal cancer (CRC) cell lines (HT-29, HCT-116 and SW-620). Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Data are presented in CRC cell lines relative to normal colorectal tissues (randomly selected 3 NATs from previous 107 cases of CRC as controls, *p<0.05, **p<0.01). NATS, non-tumor adjacent tissues.
Figure 3.
Figure 3.
The marked correlation between miR-192 and -194, miR-192 and -215, and miR-194 and -215 in colorectal cancer tissues. ΔΔCt-miRNA=(Ct-tumor-miRNA−Ct-tumor-U6RNA)−(Ct-non-tumor-iRNA−Ct-non-tumor-U6RNA). (A) Correlation in the ΔΔCt value between miR-192 and -194. (B) Correlation in the ΔΔCt value between miR-192 and -215. (C) Correlation in the ΔΔCt value between miR-194 and -215.
Figure 3.
Figure 3.
The marked correlation between miR-192 and -194, miR-192 and -215, and miR-194 and -215 in colorectal cancer tissues. ΔΔCt-miRNA=(Ct-tumor-miRNA−Ct-tumor-U6RNA)−(Ct-non-tumor-iRNA−Ct-non-tumor-U6RNA). (A) Correlation in the ΔΔCt value between miR-192 and -194. (B) Correlation in the ΔΔCt value between miR-192 and -215. (C) Correlation in the ΔΔCt value between miR-194 and -215.
Figure 3.
Figure 3.
The marked correlation between miR-192 and -194, miR-192 and -215, and miR-194 and -215 in colorectal cancer tissues. ΔΔCt-miRNA=(Ct-tumor-miRNA−Ct-tumor-U6RNA)−(Ct-non-tumor-iRNA−Ct-non-tumor-U6RNA). (A) Correlation in the ΔΔCt value between miR-192 and -194. (B) Correlation in the ΔΔCt value between miR-192 and -215. (C) Correlation in the ΔΔCt value between miR-194 and -215.
Figure 4.
Figure 4.
miR-194 significantly inhibited cell proliferation in HCT-116 cells by MTT assay (*P<0.05).
See this image and copyright information in PMC

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