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Comprehensive profiling of actionable mutations in non-small cell lung cancer (NSCLC) is vital to guide targeted therapy, thereby improving the survival rate of patients. Despite the high incidence and mortality rate of NSCLC in Vietnam, the actionable mutation profiles of Vietnamese patients have not been thoroughly examined. Here, we employed massively parallel sequencing to identify alterations in major driver genes (
EGFR
,
KRAS, NRAS, BRAF
,
ALK
and
ROS1
) in 350 Vietnamese NSCLC patients. We showed that the Vietnamese NSCLC patients exhibited mutations most frequently in
EGFR
(35.4%) and
KRAS
(22.6%), followed by
ALK
(6.6%),
ROS1
(3.1%),
BRAF
(2.3%) and
NRAS
(0.6%). Interestingly, the cohort of Vietnamese patients with advanced adenocarcinoma had higher prevalence of
EGFR
mutations than the Caucasian MSK-IMPACT cohort. Compared to the East Asian cohort, it had lower
EGFR
but higher
KRAS
mutation prevalence. We found that
KRAS
mutations were more commonly detected in male patients while
EGFR
mutations was more frequently found in female. Moreover, younger patients (<61 years) had higher genetic rearrangements in
ALK
or
ROS1
. In conclusions, our study revealed mutation profiles of 6 driver genes in the largest cohort of NSCLC patients in Vietnam to date, highlighting significant differences in mutation prevalence to other cohorts.