## 3 Results  
Comparative Analysis of the Selenium Nanorods (SeNrs) and Selenium Nanoparticles (SeNPs) Effects on the Induction of Ca\({}^{2+}\) Signals in the A-172 Human Glioblastoma Cells and L-929 Mouse Fibroblasts  
We have previously shown that spherical SeNPs induce Ca\({}^{2+}\) signaling in various cell types [24; 25]. Moreover, the amplitude of Ca\({}^{2+}\) signals and the physiological effects of SeNPs depended on the diameter of the nanoparticles [26]. The use of selenium nanorods (SeNrs) at various concentrations starting from 1 \(\upmu\)g/mL caused the generation of Ca\({}^{2+}\) signals in A-172 cells, and the increase in [Ca\({}^{2+}\)]i depended on the dose (Figure 3A). Interestingly, the rise in [Ca\({}^{2+}\)]i to the application of SeNrs occurred after a lag period of 7 \(\pm\) 1.5 min on average, and Ca\({}^{2+}\) signals were a slow growth in [Ca\({}^{2+}\)]i and its level \begin{table}
\begin{tabular}{c c c} \hline
**Gene Name** & **Forward Primer 5\({}^{\prime}\)\(\rightarrow\)3\({}^{\prime}\)** & **Reverse Primer 5\({}^{\prime}\)\(\rightarrow\)3\({}^{\prime}\)** \\ \hline CAS–3 & GCATTGAGACAGACAGTGGTG & AATAGAGATTCTTTTGTGAGCATG \\ CAS–4 & CACGCCTGGCCTCTCTCATCAT & TAGCAAATGCCCCTCAGCG \\ BAX & GGGCTGAGCATTGGCATTG & AACACACACGTCCAAGGCCTG \\ BAK & GAGACGTGCACTCATCATGGG & CAGCCACCACCCTGTGTCAATCCA \\ BCL-2 & GGTGAGACTGGGGGGGGATG & AGCCAGGAGAATCA AACAGAG \\ ATF–4 & GTGTCTCTGTGGTGTCCC & GACCCTTCTTCCCCCTTG \\ ATF–6 & AACCCTAGTGTGAGCCCTGC & GTTCAGAGCACCCTGAAGA \\ XBPu & ACTCAGACTACTGCACCCTC & GTCATAATCCCCCC \\ XBPs & CTGAGCTCCCGCAGCCGGCGCGCGC & GGTCCAAGTTGTCCAGAATG \\ GPX1 & AAGATCCAACCAAGGGGCAA & CATCAGGTCCGGGGGGGGGG \\ GPX4 & AAGATCCAACCCCAAGGCCAAG & CATCAGGTCCGGGGGGGGGG \\ TXNRD1 & CAACAAATGTTTACAAAAAAAT & ACACTGGGCTTAAACCCTG \\ TXNRD3 & CTCTTTAGAAAAGGTGATTATTATT & GCCCCACATTTTCAGACCTG \\ Nf-kB & TACTTCTCACTTTCTACTTTTGCCCCAC & GGTCTACAGAAGGCGTG \\ \hline \end{tabular}
\end{table} 
Table 1: Cont. \begin{table}
\begin{tabular}{c c c} \hline
**Gene Name** & **Forward Primer 5\({}^{\prime}\)\(\rightarrow\)3\({}^{\prime}\)** & **Reverse Primer 5\({}^{\prime}\)\(\rightarrow\)3\({}^{\prime}\)** \\ \hline GAPDH & AAGGTGGTGAAGCAGCAGCCATC & CTCTGTCCTCCTCCTCCT \\ SELENOT & TGATTGAGAACCAGTGTGATG & GGTACAACGACCTGCC \\ SELENOM & CGCCTAAGAGAGGTGAGCC & CTTGGCGTAGAAGCCGGCGC \\ SELENOF & AGGGTCCTCAGGAGAAG & CGTTCCAAG \\ SELENOK & GAAGAGGGCCCCCCAGGAAAC & GGAATTCCCCACCATGACC \\ SELENOS & GGACCAAGCCCGAGACTGTTC & CTCTTCCCATCAT \\ SELENON & AAGATGCTTCCTCTCTAGGGT & CTGTAGGGC \\ DIO2 & GCTTATCTCTGCCCCCATTG & CACATAAACGACCTCTCT \\ CHOP & CAGCTGGGGGAGCCTGA & GACCACCCTCTCTTTCC \\ GADD34 & GAGTCGCATGAAAGAGATTGTAC & ACACGCGCGCGCGCATAT \\ PUMA & TGAAGACTGTCCGCCG & GAGGACACACGCG \\ BIM & AATGGCCGGCGCGTATGATG & GCCAA  
reaching a new elevated the steady state. On the other hand, the application of spherical SeNPs to human glioblastoma A-172 cells caused a rapid growth in [Ca\({}^{2+}\)]\({}_{\text{i}}\) and also had a dose-dependent character, but the cells responded even to the appliance of 0.5 \(\upmu\)g/mL of nanoparticles (Figure 3B). Analysis of the dependence of the Ca\({}^{2+}\) signal amplitudes of A-172 cells on the NP concentration showed that EC\({}_{50}\) for SeNrs was 3.2 \(\pm\) 0.012 \(\upmu\)g/mL, and for SeNPs it was 1.9 \(\pm\) 0.007 \(\upmu\)g/mL (Figure 3E). Figure 3: Activation of the Ca\({}^{2+}\) responses of A-172 cells (**A**,**B**) and L-929 cells (**C**,**D**) by different concentrations of selenium nanorods (SeNrs) or selenium nanoparticles (SeNPs). (**E**,**F**) Dependence of the Ca\({}^{2+}\) responses amplitude of A-172 (**E**) and L-929 (**F**) cells on the different concentrations (black squares) and SeNPs (red circles). Data were approximated by a sigmoid function. The averaged Ca\({}^{2+}\) signals of cells are presented (panels (**A**–**D**)). Ca\({}^{2+}\) signals averaged over 3 independent experiments were used to plot the dose dependences of Ca\({}^{2+}\) signal amplitudes from the concentrations of nanostructures (panels (**E**,**F**)).