Screening of Recurrence Related MicroRNA in Ductal Carcinoma In Situ and Functional Study of MicroRNA-654-5p

다국어 초록 (Multilingual Abstract)

Purpose: Ductal carcinoma in situ (DCIS) contributes to 20%–30% of newly diagnosed cases of breast cancer in China. Although the breast cancer-specific mortality of DCIS is extremely low, a small proportion of DCIS patients still show relapse or metastasis, leading to poor prognosis. Little is known about the molecular mechanism for DCIS metastasis, partly due to the limited number of poor prognosis patients. This study analyzed the clinicopathological features and screened key microRNAs (miRNAs) contributing to local or distant recurrence.
Methods: The clinicopathological features of DCIS were evaluated and survival analysis were performed to clarify risk factors associated with poor prognosis. Using miRNA arrays and real-time quantitative polymerase chain reaction (RT-qPCR) on DCIS formalin-fixed and paraffin-embedded samples with or without microinvasion with different clinical outcomes, potential DCIS metastasis-related miRNAs were screened out and further validated. The influence of one identified miRNA, miRNA-654-5p, on DCIS progression was analyzed.
Results: Poor prognosis was significantly associated with larger tumor size and higher lymph node metastasis rate (both p < 0.05). Both were independent prognostic factors for DCIS. According to RT-qPCR results, distinct miRNA expression profiles were identified between DCIS and DCIS with microinvasion (DCIS-Mi) patients. In the DCIS panel, miRNA-654-5p was significantly upregulated in the patients with poor prognosis. In vitro, miRNA-654-5p promoted MDA-MB-231 cell mobility in healing tests and metastasis in the Transwell study.
Conclusion: The panel of high-risk miRNAs in DCIS and DCIS-Mi differs markedly. miRNA-654-5p is significantly upregulated DCIS patients having poor prognosis and may be essential for local and distant recurrence in DCIS.

참고문헌 (Reference)

1 Ueno K, "microRNA-183 is an oncogene targeting Dkk-3 and SMAD4 in prostate cancer" 108 : 1659-1667, 2013

2 Lu M, "miR-654-5p targets GRAP to promote proliferation, metastasis, and chemoresistance of oral squamous cell carcinoma through Ras/MAPK signaling" 37 : 381-388, 2018

3 Whipple RA, "Vimentin filaments support extension of tubulin-based microtentacles in detached breast tumor cells" 68 : 5678-5688, 2008

4 Polioudaki H, "Variable expression levels of keratin and vimentin reveal differential EMT status of circulating tumor cells and correlation with clinical characteristics and outcome of patients with metastatic breast cancer" 15 : 399-, 2015

5 O'Bryan S, "The roles of oncogenic miRNAs and their therapeutic importance in breast cancer" 72 : 1-11, 2017

6 Gregory PA, "The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1" 10 : 593-601, 2008

7 Lee RC, "The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14" 75 : 843-854, 1993

8 Östling P, "Systematic analysis of microRNAs targeting the androgen receptor in prostate cancer cells" 71 : 1956-1967, 2011

9 Roy SS, "Significance of PELP1/HDAC2/miR-200 regulatory network in EMT and metastasis of breast cancer" 33 : 3707-3716, 2014

10 Chen L, "Role of deregulated microRNAs in breast cancer progression using FFPE tissue" 8 : e54213-, 2013

11 Scribner KC, "Regulation of DCIS to invasive breast cancer progression by Singleminded-2s(SIM2s)" 32 : 2631-2639, 2013

12 Li S, "MicroRNA-132 is frequently down-regulated in ductal carcinoma in situ(DCIS)of breast and acts as a tumor suppressor by inhibiting cell proliferation" 209 : 179-183, 2013

13 Farazi TA, "MicroRNA sequence and expression analysis in breast tumors by deep sequencing" 71 : 4443-4453, 2011

14 Paydas S, "Micro-RNA(miRNA)profile in Hodgkin lymphoma : association between clinical and pathological variables" 33 : 34-, 2016

15 Tan YY, "MiR-654-5p attenuates breast cancer progression by targeting EPSTI1" 6 : 522-532, 2016

16 Rakovitch E, "Is expert breast pathology assessment necessary for the management of ductal carcinoma in situ" 87 : 265-272, 2004

17 Zha R, "Genome-wide screening identified that miR-134 acts as a metastasis suppressor by targeting integrin beta1 in hepatocellular carcinoma" 9 : e87665-, 2014

18 Hannafon BN, "Expression of microRNA and their gene targets are dysregulated in preinvasive breast cancer" 13 : R24-, 2011

19 Szynglarewicz B, "Epithelial-mesenchymal transition inducer Snail1 and invasive potential of intraductal breast cancer" 116 : 696-705, 2017

20 Mardekian SK, "Ductal carcinoma in situ of the breast : the importance of morphologic and molecular interactions" 49 : 114-123, 2016

21 Li P, "Downregulation of miRNA-141 in breast cancer cells is associated with cell migration and invasion : involvement of ANP32E targeting" 6 : 662-672, 2017

22 Lewis BP, "Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets" 120 : 15-20, 2005

23 Chen W, "Cancer statistics in China, 2015" 66 : 115-132, 2016

24 Volinia S, "Breast cancer signatures for invasiveness and prognosis defined by deep sequencing of microRNA" 109 : 3024-3029, 2012

25 Narod SA, "Breast cancer mortality after a diagnosis of ductal carcinoma in situ" 1 : 888-896, 2015

26 Fan L, "Breast cancer in a transitional society over 18 years : trends and present status in Shanghai, China" 117 : 409-416, 2009