Effects of RUNX3 mediated Notch signaling pathway on biological characteristics of colorectal cancer cells
Authors: Hang Li
Dan Li
Ning Meng
Journal: International Journal of Oncology (2017)
Discipline: CLINICAL MEDICINE
Keywords: RUNX3; Notch pathway; colorectal cancer; SW260 cells; proliferation
Affiliations: Department of General Surgery, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang 310015, P.R. China.

Abstract:  This study investigated the effects of runt-related transcription factor 3 (RUNX3) mediated Notch pathway on the biological behavior of colorectal cancer (CRC) SW260 cells. CRC tissues and para-carcinoma tissues were collected from 182 CRC patients who had undergone surgical treatment between January 2008 and December 2010. Immunohistochemical staining with streptavidin-peroxidase (SP) was used to detect RUNX3, Notch1 and Jagged 1 expression levels. CRC SW260 cells were divided into the following groups: Control group, si-NC group, si-RUNX3 group, DAPT group, si-RUNX3+DAPT group, and si-NC+DAPT group. Expression levels of RUNX3, and Notch signaling related genes were measured by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and western blotting in vitro. Besides, MTT, soft agar colony formation, Annexin V-FITC/PI double staining and Transwell were performed to analyze the effects of RUNX3 on cell growth and metastasis. Lower positive expression rate of RUNX3 and higher positive expression rate of Notch1 and Jagged 1 were observed in CRC tissues than those in normal adjacent tissues with a negative correlation, and the expression levels were associated with the differentiation degree, TNM staging, lymph node metastasis and tumor invasion depth (all P&lt;0.05). RUNX3 expression was reduced in si-RUNX3 and si-RUNX3+DAPT group but the expression levels of Notch signaling related genes were markedly increased in si-RUNX3 group or decreased in DAPT and si-NC+DAPT group, as compared with those in the control group (all P&lt;0.05). In addition, the proliferation, colony formation, migration and invasion abilities of SW260 cells were enhanced in si-RUNX3 group but were restricted in DAPT and si-NC+DAPT group, which was contrary to cell apoptosis (all P&lt;0.05). RUNX3 contributes to attenuate the proliferation and metastasis of CRC cells, and promotes cell apoptosis through inhibition of Notch signaling pathway. 

Retracted: No

PubPeer comments:
  Figure 6. Comparison of invasion and metastasis of CRC SW620 cells in each group. (A) Cell migration ability was analyzed by Transwell . ![file] The marked panels also feature in Fig 4B of [Bai et al (2016)] ... ![] ...and in Figure 13A from [Lu et al (2018)] ... and in a number of other papers from otherwise-unrelated authors, linked in that thread. ![] , 
 Fig 6, A and C. The Control panel of 6A (migration) overlaps with Control **and** si-NC panels of 6C (invasion), marked by brown and black rectangles. ![file] The si-NC panel of 6A (red rectangle) can also be seen in Fig 5B of [Xie et al (2017)] and Fig 6(a) of [Zhang et al (2019)] . ![] ![] The lower-right panel of 6C (another red rectangle, apologies for confusion) can be found in 5H of Xie et al. ![] , 
  Figure 5. Comparison of apoptosis rate and apoptosis-related protein levels of SW620 cells in each group. (A) Flow cytometry was conducted to measure SW620 cell apoptosis in each group . ![file] Resolution is not high. Still, it is enough. ![file] ,