Through this work, green tea extract—an environmentally friendly reducing and capping agent—has been used to manufacture titanium dioxide nanoparticles (TiO2.NPs). Therefore, we investigated in this work how TIO2 nanoparticles could be harmful to the growth of HuH-7 cells (human Hepatocellular cancer cell line). The cytotoxic effect and the UV–Vis (ultraviolet–visible) spectra of TIO2 (NPs) have been investigated following exposure to a 250W infrared radiation lamp for varied durations, ranging from 2.5 to 20min. The MTT test was used to measure the number of viable cells, which was used to assess the cytotoxic effect. The survival curve of each tumour cell line following treatment with the designated substance is obtained by plotting the relationship between remaining cells and drug concentration. Using visual representations of the dose response curve for each conc., the 50% inhibitory concentration (IC50), or the dosage needed to produce harmful effects in 50% of undamaged cells, was calculated. According to the current analysis, infrared radiation (IR) is treated as thermal radiation and has the same effect as annealing temperature. As a result, the IC50 values of HuH-7cells for the 5-FU Standard, the control (TIO2.NPs prior to irradiation) and TIO2.NPs following exposure to 20m IR-irradiation, respectively, were 12.8.8±3.2 μg/m, 60.43±7.9 μg/mL, and 80.1±21.3 μg/mL. The IC50 values for the 5-FU Standard, TIO2.NPs before irradiation, and TIO2.NPs after exposure to 20m IR-irradiation were 65,1±9.8 μg/mL, 64,7±12.48 μg/mL, and 98,6±20.2 μg/mL, respectively, in normal WI38 cells.