Isolating the genes responsible for cadmium (Cd) accumulation and tolerance in oilseed rape and uncovering their functional mechanism is of great significance for guiding genetic improvement to cope with heavy metal pollution. In this study, we screened the cDNA library of Brassica napus cv. Westar using a yeast genetic complementation system and isolated BnMT2-22a, BnMT2-22b and BnMT3b, which can mediate Cd tolerance in yeast. They all have two cysteine-rich domains in their sequence. Ectopic expression of these MTs demonstrated that all of them enhanced Cd and Cu tolerance in yeast, but had no effect on Mn and Zn tolerance. The fusion of the red fluorescent protein mRFP did not affect their function in mediating Cd tolerance, and using these functional fusion proteins we observed that they were all localized in cytosol. Meanwhile, their expression in yeast did not affect the accumulation of Cd in the yeast transformants. Gene expression analyses found that BnMT2-22a, BnMT2-22b and BnMT3b were all induced by Cd in roots, and BnMT3b was also significantly induced in shoots. These results indicate that the genes BnMT2-22a, BnMT2-22b and BnMT3b isolated with cDNA library screening can mediate Cd tolerance, and they may detoxify Cd via cytosolic chelation.

Breast cancer represents a persistent global health burden, marked by extensive molecular heterogeneity and frequent therapeutic resistance in aggressive subtypes, particularly triple-negative breast cancer (TNBC). These clinical challenges underscore the urgency for alternative therapeutic strategies. Bioactive alkaloids isolated from Nelumbo nucifera, especially the bisbenzylisoquinoline compounds liensinine (LIE), isoliensinine (ISO), and neferine (NEF), have emerged as promising candidates due to their ability to disrupt oncogenic signaling pathways and inhibit malignant cellular transformation. The present study conducted a systematic investigation of LIE, ISO, and NEF across multiple breast cancer cell lines, including highly aggressive TNBC models. Results revealed potent growth-inhibitory effects mediated through apoptosis induction and cell cycle arrest at both the G1 and G2/M phases. Furthermore, transcriptomic profiling and molecular analysis identified LIE as a principal effector, driving extensive transcriptional reprogramming and targeting the MAPK and mTOR pathways as core regulators of its anti-cancer efficacy. Collectively, these findings define a mechanistic framework for the anti-cancer potential of N. nucifera-derived alkaloids and provide a compelling foundation for their development as therapeutic candidates for advanced breast cancer.