BACKGROUND: Conventional hamstring (HAM) stretching therapeutic effects are not substantiable in neuromusculoskeletal conditions with HAM tightness or shortness. We developed a kinetic chain stabilization exercise to provide a more sustainable effectiveness in adults with HAM tightness. However, its therapeutic effects and underlying motor mechanisms remain unknown. OBJECTIVE: To compare the effects of traditional active HAM stretching (AHS) and kinetic chain stretching (KCS) on electromyographic (EMG) amplitude and hip flexion range of motion (ROM) in participants with HAM tightness. METHODS: In this randomized controlled trial, 18 participants (mean age: 25.01 ± 2.47 years) with HAM tightness were assigned to the AHS or KCS group. Hip joint movement, EMG amplitude, and onset times were recorded in the bilateral erector spinae, HAM, transverse abdominis/internal oblique (IO), external oblique (EO), and rectus abdominis during a straight leg raise test. RESULTS: Compared to AHS, KCS led to greater increase in the hip flexion ROM and EMG activation amplitudes in the left and right EO and left IO. Post-test hip flexion ROM data in both the groups were higher than the pre-test data. CONCLUSION: KCS produced more sustainable effectiveness in hip flexion movement and EMG motor control patterns in participants with HAM tightness than AHS.