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. 2017 Oct;14(4):4619-4624.
doi: 10.3892/ol.2017.6730. Epub 2017 Aug 7.

Role and mechanism of action of LRIG1 in ovarian cancer cell line and VP16 drug-resistant cell line

Yaqi Zhang  1 , Zhizhen Liu  1 , Shunrui Yu  1
Affiliations

Role and mechanism of action of LRIG1 in ovarian cancer cell line and VP16 drug-resistant cell line

Yaqi Zhang et al. Oncol Lett. 2017 Oct.

Abstract

We investigated the role of leucine-rich repeats and immunoglobulin-like domains (LRIG)-1 in ovarian cancer cell line and VP16 drug-resistant cell line to explore the possible mechanism of action. Human ovarian cancer cell line SKOV3 and the VP16 drug-resistant cell line SKOV3/VP16 were used to investigate whether LRIG1 affects the sensitivity of SKOV3 to drugs. RT-qPCR was used to detect the difference in LRIG1 expression between drug-resistant and wild-type cell lines. siRNA LRIG1 was designed and transfected to silence LRIG1 to investigate the mechanism by which LRIG1 affects the sensitivity of SKOV3 to drugs. Wild-type cells were transfected with SKOV3. The cells were divided into 3 groups (VP16, NC + VP16 and siRNA LRIG1 + VP16 treatment group). VP16 (IC50 value) was added 24 h after transfection. The CCK-8 method was used to detect the proliferation of each group at multiple time points (0, 24, 48 and 72 h). A colony-forming assay was used to detect cell proliferation and flow cytometry was used to detect cell apoptosis. The expression of LRIG1 was lower in the drug resistant cell line than that of the wild-type cell line. The expression of LRIG1 significantly decreased with the increase of VP16 concentration (P<0.05). The apoptotic rate was decreased but there was an increase on cell clones in the siLRIG1 + VP16-treated group as compared to VP16- and NC+ VP16-treated groups (P<0.05). The LRIG1 gene affects the sensitivity of SKOV3 cells to drug in a dose-related manner, indicating that the reduced expression of LRIG1 can inhibit cell apoptosis.

Keywords: SKO; VP16; cancer; drug resistance; leucine-rich repeats and immunoglobulin-like domains 1; ovarian.

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Figures

Figure 1.
Figure 1.
Expression of LRIG1 detected by RT-qPCR, (A) Compared with wild-type cells, LRIG1 expression in drug-resistant cells was significantly reduced (P<0.05). (B) The expression of LRIG1 was decreased significantly with the increase of VP16 concentration (*P<0.05). LRIG1, leucine-rich repeats and immunoglobulin-like domains.
Figure 2.
Figure 2.
CCK-8 to detect tumor cell inhibition rates at different concentrations of VP16.
Figure 3.
Figure 3.
The expression levels of LRIG1 in SKOV3 cells after siLRIG1 transfection. Western blot analysis showed that the expression of LRIG1 protein in the siRNA LRIG1 + VP16-treated group was significantly lower than that in the VP and the NC+VP16-treated group (*P<0.05). LRIG, leucine-rich repeats and immunoglobulin like domains.
Figure 4.
Figure 4.
Cell viability detected by CCK-8 method. The cell viability of siRNA LRIG1 + VP16 treatment group was significantly higher than that of VP16 and NC + VP16, suggesting that silencing LRIG1 can promote cell viability. LRIG1, leucine-rich repeats and immunoglobulin-like domains; CCK, cell counting kit.
Figure 5.
Figure 5.
Cell apoptosis detected by flow cytometry. Compared with VP16 and NC + VP16 treatment group, the proportion of apoptotic cells in siLRIG1 + VP16 treatment group was significantly decreased (P<0.05).
Figure 6.
Figure 6.
Cell proliferation detected by colony formation assay. (A) Reprentative results of the colony formation assay of cells in VP16, NC + VP16 treatment group and siRNA LRIG1 + VP16 treatment group. (B) Compared with VP16 and NC + VP16 treatment group, the number of colonies formed by cells in the siRNA LRIG1 + VP16 treatment group was increased significantly (*P<0.05).
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