The invention provides a cell-penetrating peptide with negative charge and its homologues, having a cell internalization function. Net charge of the cell-penetrating peptide under physiological conditions is a negative value; an amino acid sequence is ELDGDV, LQEDTPP, DPNEKRD, DHMKQHD, NYTWDQW, TTDRHDL, SEQPAHS, DHYHPFS, SDAKNDL, DPYHPFS, SLTNTEH, TVSKGEE, EHYHPFM, TDTTPLQ, GGSAETG, DGPGTAA, SLTAPDY, PLDVNNM, DGTPVGM, LQNLGEF, DSHLSLM, VGSPDGL, DLNLPFA or PLLEGSL. The invention also provides application as an intracellular transport carrier and application of the cell-penetrating peptide withnegative charge as therapeutic drugs in terms of pharmaceutical application and imaging of the intracellular transport carrier.