BACKGROUND: WNT inducible secreted protein 2 (WISP2) has been linked with a variety of human cancer types and may contribute to cancer metastasis. The current study investigated the importance of WISP2 in colorectal cancer cells, examining the impact of targeting WISP2 on Caco-2 cell invasion and motility together with potential mechanisms of action. MATERIALS AND METHODS: WISP2 expression was targeted in Caco-2 cells using a ribozyme transgene system and successful knockdown was verified using reverse transcription-polymerase chain reaction (RT-PCR). The impact of WISP2 knockout (Caco-2(WISP2 KO)) on cell growth, adhesion, motility and invasion was examined using a number of in vitro functional assays. In vitro invasion assays were repeated in the presence of wingless-type MMTV integration site family (WNT) inhibitors (FH535 and IWP-2) to investigate the role of the WNT-signalling pathway in the regulation of cell invasion by WISP2. Quantitative-PCR was conducted to measure matrix metalloproteinase (MMP) expression in control [wild-type (Caco-2(WT)) and cells containing the empty pEF6 plasmid (Caco-2(pEF6))] and Caco-2(WISP2 KO) cells. RESULTS: WISP2 knockout resulted in a significant increase in Caco-2 cell invasion and motility (p<0.05 in comparison to wild-type and plasmid control Caco-2 cells). WISP2 knockout had no significant effect on Caco-2 cell growth rate in 3- and 5-day incubation and no significant impact on Caco-2 cell-matrix adhesion rates (p>0.05). Expression analysis of a number of MMPs indicated an insignificant up-regulation of MMP2, MMP9 (p>0.05) but significant up-regulation of MMP7 (p=0.025) in Caco-2(WISP2 KO) cells compared to controls. Inhibition of WNT signalling using FH535 and IWP-2 brought about a significant or borderline significant decrease in Caco-2(WISP2 KO) cell invasion (FH535 p=0.065) and (IWP-2 p=0.002) and negated the pro-invasive effect of targeting WISP2 in Caco-2 cells. CONCLUSION: WISP2 knockout significantly increased Caco-2 cell invasion and motility. Up-regulation of MMP2, -7 and -9 may indicate that WISP2 regulates invasion and motility through MMPs. Regulation of invasion by WISP2 may involve the WNT signalling pathway.
BACKGROUND:WNT inducible secreted protein 2 (WISP2) has been linked with a variety of humancancer types and may contribute to cancer metastasis. The current study investigated the importance of WISP2 in colorectal cancer cells, examining the impact of targeting WISP2 on Caco-2 cell invasion and motility together with potential mechanisms of action. MATERIALS AND METHODS:WISP2 expression was targeted in Caco-2 cells using a ribozyme transgene system and successful knockdown was verified using reverse transcription-polymerase chain reaction (RT-PCR). The impact of WISP2 knockout (Caco-2(WISP2 KO)) on cell growth, adhesion, motility and invasion was examined using a number of in vitro functional assays. In vitro invasion assays were repeated in the presence of wingless-type MMTV integration site family (WNT) inhibitors (FH535 and IWP-2) to investigate the role of the WNT-signalling pathway in the regulation of cell invasion by WISP2. Quantitative-PCR was conducted to measure matrix metalloproteinase (MMP) expression in control [wild-type (Caco-2(WT)) and cells containing the empty pEF6 plasmid (Caco-2(pEF6))] and Caco-2(WISP2 KO) cells. RESULTS:WISP2 knockout resulted in a significant increase in Caco-2 cell invasion and motility (p<0.05 in comparison to wild-type and plasmid control Caco-2 cells). WISP2 knockout had no significant effect on Caco-2 cell growth rate in 3- and 5-day incubation and no significant impact on Caco-2 cell-matrix adhesion rates (p>0.05). Expression analysis of a number of MMPs indicated an insignificant up-regulation of MMP2, MMP9 (p>0.05) but significant up-regulation of MMP7 (p=0.025) in Caco-2(WISP2 KO) cells compared to controls. Inhibition of WNT signalling using FH535 and IWP-2 brought about a significant or borderline significant decrease in Caco-2(WISP2 KO) cell invasion (FH535 p=0.065) and (IWP-2 p=0.002) and negated the pro-invasive effect of targeting WISP2 in Caco-2 cells. CONCLUSION:WISP2 knockout significantly increased Caco-2 cell invasion and motility. Up-regulation of MMP2, -7 and -9 may indicate that WISP2 regulates invasion and motility through MMPs. Regulation of invasion by WISP2 may involve the WNT signalling pathway.
Entities:
Keywords:
WISP2; cell invasion; cell motility; colorectal cancer
Authors: Thomas J Lynch; Preston J Anderson; Weiliang Xie; Adrianne K Crooke; Xiaoming Liu; Scott R Tyler; Meihui Luo; David M Kusner; Yulong Zhang; Traci Neff; Daniel C Burnette; Katherine S Walters; Michael J Goodheart; Kalpaj R Parekh; John F Engelhardt Journal: Stem Cells Date: 2016-07-11 Impact factor: 6.277
Authors: Cynthia Yu-Wai-Man; Nicholas Owen; Jonathan Lees; Aristides D Tagalakis; Stephen L Hart; Andrew R Webster; Christine A Orengo; Peng T Khaw Journal: Sci Rep Date: 2017-07-17 Impact factor: 4.379
Authors: Benjamin A Neely; Christopher E Wilkins; Laura A Marlow; Dariya Malyarenko; Yunee Kim; Alexandr Ignatchenko; Heather Sasinowska; Maciek Sasinowski; Julius O Nyalwidhe; Thomas Kislinger; John A Copland; Richard R Drake Journal: PLoS One Date: 2016-04-29 Impact factor: 3.240
Authors: Sebastian Schölch; Sebastián A García; Naoki Iwata; Thomas Niemietz; Alexander M Betzler; Lahiri K Nanduri; Ulrich Bork; Christoph Kahlert; May-Linn Thepkaysone; Anka Swiersy; Markus W Büchler; Christoph Reissfelder; Jürgen Weitz; Nuh N Rahbari Journal: Oncotarget Date: 2016-05-10