The Sue Ryder store in Queen Street, Whittlesey, has shut after its lease came to an end.
The store shut on Friday, March 20.
The nearest Sue Ryder charity shop is now in Whittlesey Road, Stanground.
Meta Lear, Sue Ryder area manager, said the Whittlesey store required more funding which would divert money away from vital services.
She added: “It is with much regret that we took the decision not to renew the lease.
“We have enjoyed being a part of the town’s community and are very sad to leave.
“We have done everything we possibly can to offer alternative employment for the shop’s staff and are pleased that all of them will remain working for Sue Ryder.
“We would like to thank them and the volunteers who have supported them for all their outstanding work and support to the charity.”