**2.7 LncRNA-based prognostic signature optimized with lasso regression for ovarian cancers**

Lasso regression means least absolute shrinkage and selection operator regression, which was used to optimize and construct lncRNA-based prognostic signature, and the glmnet R package was used to measure the association between survival risk and lncRNA signature in ovarian cancers. Moreover, univariate and multivariate Cox regression, and Kaplan–Meier method were used to identify overall survivalrelated clinical characteristics described above in ovarian cancers to confirm the established lncRNA-based prognostic model. The detailed procedure was described previously [4].

and 22.54 μM for TOV-21G (**Figure 1A**). The IC50 of ovarian cancers were significantly lower than the normal controls. Further, 20 μM ivermectin - slightly lower than IC50 – can effectively inhibit ovarian cancer proliferation (**Figure 1B** and **C**) [21]. For *in vivo* human trial, the highest FDA-approved ivermectin dose was 200 μg/kg for human use in anti-parasite; however, a study on 68 human subjects found that the dose up to 2,000 μg/kg still worked well without CNS toxicity. The mean area under the curve ratios for the 30 and 60 mg doses were 1.24 and 1.40, indicating a minimal accumulation of ivermectin [5, 22]. These data demonstrate that ivermectin was a well-tolerated safe drug. Second, EdU cell proliferation experiments also confirmed that ivermectin significantly suppressed cell proliferation of ovarian cancers with a time-dependent relationship (**Figure 1D**-**F**) [21]. Third, Clonogenic survival experiments confirmed that ivermectin effectively inhibited the formation of cell clones with a time-dependent relationship (**Figure 1G**-**H**) [21]. Moreover, 10 μM ivermectin cannot effectively inhibit cell proliferation of ovarian cancers, 30 μM ivermectin caused cell death of ovarian cancers, and 20 μM ivermectin was a suitable dose to significantly suppress growth

*Ivermectin suppressed ovarian cancer cell proliferation* in vitro*, measured with CCK8 (A-C), EdU (D-F), and clonogenic experiments (G, H). Reproduced from Li et al. [21], with copyright permission from nature springer*

*The Anti-Cancer Effects of Anti-Parasite Drug Ivermectin in Ovarian Cancer*

*DOI: http://dx.doi.org/10.5772/intechopen.95556*

**3.2 Effects of ivermectin on cell cycle and apoptosis in ovarian cancers**

Flow cytometry was used to measure cell cycle and apoptosis of ovarian cancer cells treated with and without ivermectin (**Figure 2**) [21]. First, the cell proportion was significantly increased in G0/G phase, decreased in S phase, and no change in G2/M phase in the high concentration (20- and 30-μM) compared to the low concentration (0- and 10-μM) of ivermectin groups (**Figure 2A**-**C**). Second, compared to control group, the proportion of apoptosis cells was significantly increased

and proliferation of ovarian cancer cells.

**Figure 1.**

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*publisher, copyright 2020.*

## **2.8 Statistical significance**

Benjamini–Hochberg (FDR) for multiple testing was used to correct the p values of IPA, GO, and KEGG analyses. Student's t test was used for qRT-PCR and western blot data (p < 0.05) with data expression of mean SD (n = 3).
