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In vivo, \u03b2-aminopropionitrile-mediated tumor softening loosened ECM architecture, increased NP penetration, and promoted cellular uptake by reducing F-actin-associated mechanical resistance. Intratumoral spatial analyses and paired bilateral tumor experiments further supported local regulation of NP uptake by the actomyosin state under matched matrix conditions. Combined tumor softening and donafenib-loaded PLGA NPs reduced endpoint tumor weight by 87.8% relative to free donafenib but increased pulmonary metastasis, revealing a potential efficacy-safety trade-off. 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