CD133 Affects the Invasive Ability of HCT116 Cells by Regulating TIMP-2

Abstract

CD133 is widely expressed in colorectal cancer (CRC) tissues and cell lines. This protein has been used as a marker of CRC cancer stem cells, although the function and mechanism of CD133 in CRC invasion and metastasis remain unclear. In our study, we examined the role of CD133 in CRC invasion in vitro and investigated the mechanism involved in CD133-related invasion. CD133(high) and CD133(low) HCT116 cells were isolated, and the proliferation and invasive ability of these two subpopulations were tested. CD133(high) HCT116 cells exhibited greater proliferation and invasion compared with CD133(low) HCT116 cells. CD133 knockdown (using CD133 small-interfering [si]RNA) inhibited the invasive activity of CD133si-HCT116 cells. For the first time, we found that the expression of tissue inhibitor of matrix metalloproteinases-2 (TIMP-2) was down-regulated in CD133si-HCT116 cells. In addition, for the TIMP-2si-HCT116 cells (transfected with TIMP-2 siRNA), in vitro invasion was significantly decreased, whereas the expression of CD133 remained unchanged. Finally, the metalloproteinase 2 and MEK/ERK signaling pathways were examined, and no significant change was observed after the knockdown of CD133 or TIMP-2 in HCT116 cells. In conclusion, we demonstrated that CD133 plays an important role in HCT116 cell invasion, and for the first time, we found that CD133 knockdown significantly down-regulated TIMP-2 expression, which suggests that CD133 affects the invasive ability of HCT116 cells by regulating TIMP-2.

NEDD9 Depletion Leads to MMP14 Inactivation by TIMP2 and Prevents Invasion and Metastasis

“…TIMP2 is a potent inhibitor of most soluble MMPs (39), thus TIMP2 depletion could also relieve the inhibition of other soluble MMPs produced by cells. Nevertheless, our findings indicate that depletion of TIMP2 alone is not sufficient to restore the activity of soluble MMPs most likely due to the presence of other TIMPs such as TIMP1, 3 and 4 (39–40). Re-expression of NEDD9 restores the MMPs activity and invasion proficiency of shNEDD9 cells.…”

The Role of Matrix Metalloproteinases in Colorectal Cancer

“…In Korean CRC patients, a genetic polymorphism in TIMP-2 was associated with increased risk of metastasis and worse prognosis [40]. Colon cancer cells with siRNA knockdown of CD133, a putative stem cell and cancer stem cell marker, displayed down-regulated TIMP-2 expression and decreased invasiveness [41]. …”

CD133: a stem cell biomarker and beyond

“…In colorectal cancer with early liver metastases, co-expression of CD133 and CD44 is significantly higher when compared to those without early liver metastases [48]. Knockdown of CD133 in hepatocarcinoma PLC/PRF/5 and HCT116 cells results in decreased expressions of matrix metalloproteinase (MMP)-2, a disintegrin and metalloproteinase (ADAM)9 [55,56]. These lead to decreased invasion as demonstrated in an in vitro system [55,56].…”

“…Knockdown of CD133 in hepatocarcinoma PLC/PRF/5 and HCT116 cells results in decreased expressions of matrix metalloproteinase (MMP)-2, a disintegrin and metalloproteinase (ADAM)9 [55,56]. These lead to decreased invasion as demonstrated in an in vitro system [55,56]. In addition, chemokine CCL5 and its receptors, CCR1, CCR3 and CCR5, are found to be upregulated in CD133 + cancer stem-like cells from ovarian cancer [57].…”