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Breast cancer is one of the most virulent types of cancers that contribute to high mortality worldwide. Hyperthermia (HT) therapy was introduced as one of the alternatives to treat breast cancer. However, HT caused poor specific heat distribution in tumor during treatment which lead to unintentional destruction of normal cells. Therefore, this study explores the integration effect of multiwalled-carbon nanotubes (MWCNT) in combination with hyperthermia for breast cancer therapy regime. MWCNT was recognized as the potential candidate for hyperthermal therapy due to its thermal conductance and stability which generated a significant amount of heat during hyperthermia. This subsequently will provide specific heat distribution in tumor, which expected to improve the treatment outcome. In this study, 5 × 105 EMT6 breast cell was inoculated subcutaneously into right flank of female Balb/c mice. At day 7 post-inoculation, tumors were intratumorally injected with MWCNT and subjected to local HT for 3 consecutive days. The tumor progression was monitored every 2 days. In another experiment, mice were sacrificed after 24 hr post-treatment and tumors were harvested for pathological examination by H&E and immunohistochemical analysis for PCNA and Hsp70 expression. Results from this study demonstrated that mice from combined treatment displayed complete tumor eradication and significantly prolonged median survival (P<0.001). Furthermore, histological analysis of tissues from combined treatment showed that the tumor experienced cell necrosis. For Hsp70 expression, HT and combined treatment both showed expression of Hsp70 protein corresponding to hyperthermia. In addition, significant reduction of PCNA-positive cells was observed in treated group indicated that HT in combination with MWCNT treatment inhibited tumor cells proliferation. Altogether, results presented in this study suggested that MWCNT might have a potential as an anticancer therapeutic agent in future breast cancer treatment.